TW201021828A - Antibodies and related molecules that bind to 58P1D12 proteins - Google Patents

Abstract

Antibodies and molecules derived therefrom that bind to 58P1D12 protein and variants thereof, are described wherein 58P1D12 exhibits tissue specific expression in normal adult tissue, and is aberrantly expressed in the cancers listed in Table I. Consequently, 58P1D12 provides a diagnostic, prognostic, prophylactic and/or therapeutic target for cancer. The 58P1D12 gene or fragment thereof, or its encoded protein, or variants thereof, or a fragment thereof, can be used to elicit a humoral or cellular immune response; antibodies or T cells reactive with 58P1D12 can be used in active or passive immunization.

Description

201021828 VI. OBJECTS OF THE INVENTION: TECHNICAL FIELD The invention described in this patent specification relates to antibodies and binding fragments thereof which can bind to a protein named 58P1D12 and molecules engineered therefrom. The invention further relates to diagnostic, prognostic, prophylactic and therapeutic methods and compositions suitable for the treatment of cancers exhibiting 58P1D12. RELATED APPLICATIONS This application claims US Provisional Patent Application No. 097131869, filed on August 20, 2008, and Taiwan Application No. 098107574, filed on March 20, 2008, filed on August 20, 2008 The priority of U.S. Provisional Application No. 61/153, 225, filed on Jan. 20, 2008, and U.S. Provisional Application No. 61/153,225, filed on Jan. 20, 2008. The contents of these applications are hereby incorporated by reference in their entirety. References to the Sequence Listing submitted via the EFS website are as authorized and set forth in MPEP §1730 II.B.2(a)(C), and the entire contents of the Sequence Listing via the USPTO EFS Website Server are referenced below. All are incorporated herein for all purposes. The sequence listing of the following text files submitted electronically has been identified: File name creation period size (bytes) 511582002083Seqlist.txt July 14, 2009, 57, 186 [Prior Art] Cancer is the second major cause of human death, Second only to coronary artery disease. Millions of people die of cancer every year around the world. According to the American Cancer Society, the number of cancer deaths in the United States alone exceeds 500,000 a year, with more than 1.2 million new diagnoses per year. Although the deaths from heart disease have decreased significantly, the deaths from cancer have generally increased. In the early part of the next century, cancer is expected to be the leading cause of death. = All = within the bounds, there are several types of cancer that attract attention as the main killer. In particular, 'lung cancer, prostate cancer, breast cancer, colon cancer, pancreatic cancer, nest cancer, and bladder cancer represent the leading cause of cancer death. These and almost all other cancers share a common fatal feature. With a few exceptions, cancer metastatic disease is fatal. Moreover, even for those cancer patients who were initially free from their primary cancer, the common experience has shown a significant change in their longevity. Many cancer patients experience severe anxiety because they know the likelihood of recurrence or treatment failure. Many cancer patients experience physical weakness after treatment. In addition, many cancer patients experience relapse. Prostate cancer is the fourth most prevalent cancer in men worldwide. In North America and Northern Europe, it is by far the most common cancer in men and the second leading cause of cancer death in men. In the United States alone, the annual mortality of this disease is much steeper than 3,000 people, second only to lung cancer. Although these numbers have a f-threshold 35, there is no effective treatment for metastatic prostate cancer. Surgery 】! Abdominal surgery, radiation therapy, hormone excision therapy, surgical cutting and chemotherapy are still the main forms of treatment. Unfortunately, these treatments are ineffective for many people and often have adverse consequences. Ί 7 刖, lack of accurate detection of early localized tumors of the prostate tumor 仏 ~ still clear limitations in the diagnosis and control of this disease. Although serum 142769.doc 201021828 has been a very useful tool for the gland-specific antigen (PSA) assay, it is recognized to lack specificity and general utility in several important aspects. Advances in the identification of other specific markers for prostate cancer have been improved by the generation of prostate cancer xenografts that can reproduce the different stages of the disease in mice. LAPC (Loss Angeles prostate cancer) xenografts are prostates that have survived in severely complex immunodeficiency (SCID) mice and have demonstrated the ability to mimic androgen-dependent conversion to androgen-independent Cancer xenografts (Klein et al., 1997, Nat. Med. Ref. 3:402). Recently identified prostate cancer markers, including pcTA-ysu et al, 1996, Proc. Natl. Acad. Sci. USA 93: 7252), prostate specific membrane (PSM) antigens (Pinto et al, Clin Cancer Res, September 1996) 2 days

(9): 1445-51), STEAP (Hubert et al, Proc Natl Acad Sci U S A. December 7, 1999; 96(25): 14523-8) and prostate stem cell resistance

Original (PSCA) (Reiter et al. '1998, Proc. Natl. Acad. Sci. USA 95: 1735) 〇$ Although previously identified targets (such as PSA, PSM, PCTA, and 5 8P1D12) are helpful in diagnosing and treating the prostate Cancer, but for further improvement in diagnosis and treatment, it is still necessary to identify other targets and therapeutic targets for prostate cancer and related cancers. Renal cell carcinoma (RCC) accounts for approximately 3% of adult malignancies. Once the adenoma reaches a diameter of 2 to 3 cm, there is a possibility of malignancy. In adults, the two major malignant renal tumors are renal cell adenocarcinoma and transitional cell carcinoma of the renal pelvis or ureter. In the United States, the incidence of renal cell adenocarcinoma is estimated to exceed 29, and in 1998 more than 11,600 patients died of this disease. Migration is fine 142769.doc 201021828 Cell carcinoma is less common and occurs in the United States at an annual rate of about 5 cases. Surgery has been the primary treatment for renal cell adenocarcinoma for decades. Until recently, any systemic therapy still failed to cure metastatic disease. With the recent developments in systemic therapies, particularly immunotherapy, it is possible to invasively approach metastatic renal cell carcinoma of a suitable patient with a promising response. Despite this, there is still a need for effective therapies for such patients. Among all new cancer cases in the United States, bladder cancer accounts for the majority of men (the fifth most common tumor) and 3% of women (the eighth most common tumor). As the elderly population increases, the incidence rate rises slowly. In 998, an estimated 54,500 cases were included, including 39 5 males and 15 females. In the United States, the age-adjusted incidence rate is “for every 〇〇, and for every 〇〇, for every 〇〇, 8 for women. The male/female 3: i historical ratio may continue to decrease, This is related to the female smoking pattern. In 1998, it was estimated that bladder cancer caused 4000 deaths (7,800 males and 39 females). The incidence and mortality of bladder cancer increased sharply with age and became older as people became older. It becomes a problem. Most bladder cancers recur in the bladder. Bladder cancer is controlled by a combination of transurethral cystectomy (TUR) and intravesical chemotherapy or immunotherapy. The multifocal and recurrent nature of bladder cancer illustrates the limitations of TURi. Most muscle invasive cancers cannot be cured by themselves. Radical bladder dissection and urinary diversion are the most effective ways to eliminate cancer, but have an undeniable effect on urinary function and sexual function. For patients with bladder cancer. There is still a great need for the form of treatment. In 2000, there were an estimated 13 cases of colorectal cancer in the United States. 142769.doc 201021828 includes 93, _ cases of colon cancer and 3M cases of rectal cancer Cases. Colorectal cancer is the third most common cancer in men and women. During 1992-1996, the incidence was significantly reduced (21% per year). Studies have shown that this decrease is due to increased 4 tests and polyp removal. To prevent the development of polyps into invasive cancer. In 2000, there were an estimated 56,3 deaths (1), 7 7 deaths from colon cancer, 8 deaths from rectal cancer, accounting for the total number of cancer deaths in the United States. 11%. φ Vision, surgery is the most common form of treatment for colorectal cancer, and it can usually be cured for cancers that have not spread. For cancer patients who have penetrated the intestinal wall or have spread to most of the lymph nodes, Before or after surgery, you can order chemotherapy or chemotherapy plus radiation. Colon cancer sometimes requires permanent colostomy (formation of abdominal openings to eliminate human waste) and rectal cancer is occasionally needed. Effective diagnosis and treatment of colorectal cancer Forms still exist. In 2000, there were an estimated 164,1 new cases of lung and bronchial cancer, φ accounting for 4% of all cancer diagnoses in the United States. Lung and bronchial cancer The incidence rate in sex has been significantly reduced, from 10 to 10 in 1984, and as high as 86 5 to 70.0 in 1996. In the 1990s, the growth rate among women began to slow down. In 1996, women The incidence rate was 42 3 per 1〇〇〇〇〇. In 2000, lung and bronchial cancer were estimated to cause 156,9 deaths, accounting for 28% of all cancer deaths. During 1992-1996, lung cancer died. The rate is significantly lower among men (17% per year), while female mortality is still rising significantly (up 9% per year). Since 1987, more women die of lung cancer each year than women who die from breast cancer. 4 years of female cancer death for many years J42769.doc 201021828 The main reason. The reduction in lung cancer morbidity and mortality is most likely due to the decline in smoking rates in the past three years; however, the decline in women's smoking patterns lags behind that of men. Worrying, although the decline in adult (4) consumption has slowed, the tobacco consumption of young people has risen again. Treatment options for lung and bronchial cancer are based on the type and stage of cancer and include surgery, radiation therapy, and chemotherapy. For a variety of localized cancers, surgery is generally chosen. Since the disease has generally spread to the time of discovery, it is often necessary to combine radiation therapy and chemotherapy with surgery. Chemotherapy, alone or in combination with radiation, is selected for the treatment of small cell lung cancer; most patients experience remission after treatment with this therapy, and in some cases, this relief is long-lasting. However, there is a continuing need for effective treatment and diagnostic methods for lung and bronchial cancer. It is estimated that during the year 2000, there will be an estimated 182,8 new cases of invasive breast cancer among women in the United States. In addition, approximately 1,400 new cases of breast cancer are expected to be diagnosed in men in 2000. The incidence of breast cancer in women increased by about 4% per year in the 1980s and stabilized to about 110.6 per 100 people in the 1990s. In the United States alone, it is estimated that in 2001, 41,2 (a female, 4, and 8 males) died of breast cancer. Breast cancer ranks second among female cancer deaths. Skirts According to the latest data, between 1992 and 1996, the mortality rate was significantly reduced, &# white and black young women fell the most. These reductions are mostly the result of early detection and treatment improvement. Considering the medical environment and patient preferences, breast cancer treatment may include lumpectomy (local removal of the tumor) and axillary lymph node removal; mastectomy (surgery 142769.doc 201021828 removal of the breast) and axillary lymph node removal; radiation therapy Chemotherapy; or hormone therapy. Usually, two or more methods are used in combination. Many studies have demonstrated that long-term survival after early disease's lumpectomy plus radiation therapy is similar to survival after modified radical mastectomy. Significant advances in reconstruction techniques offer several options for breast reconstruction after mastectomy. Recently, this reconstruction has been performed simultaneously with mastectomy.

Local resection of breast ductal carcinoma in situ (DCIS) with sufficient surrounding normal breast tissue prevents local recurrence of DCIS. Breast radiation and/or tamoxifen reduce the incidence of DCIS in the remaining breast tissue. This is important because if left untreated, DCIS develops into invasive breast cancer. Despite this, there are still serious side effects or continuations in these treatments. Therefore, effective breast cancer treatment is needed. In 2000, it was estimated that there were 23,1 cases of new ovarian cancer in the United States. It accounts for 4% of all cancer in women and ranks second among gynecological cancers. During 1992-1996, the incidence of India's nest cancer was significantly reduced. In 2 years, it is estimated that 14 cases of death due to nest cancer. Nested cancer causes more deaths than any other cancer in the female reproductive system. Surgery, radiation therapy and chemotherapy are the treatment options for Indian cancer. Surgery - generally involves the removal of one or two nests, the printing tube (transprint tube - nesting resection) and the uterus (hysterectomy in some very early stage tumors, only removing the nests involved, Especially for young women with good shirts. In the advanced disease towel, 'try to remove all intra-abdominal diseases to enhance the role of chemotherapy. There is still a strong need for effective treatment options for (4) nest cancer. 142769.doc 201021828 In 2000, estimated There are 28, 3 cases of new pancreatic cancer in the United States. In the past 20 years, the incidence of pancreatic cancer in men has decreased. The incidence of pancreatic cancer in women has remained roughly constant and may begin to decrease. In 2 years, in The United States estimates that pancreatic cancer causes 28,200 deaths. In the past two years, there has been a small but significant decrease in male mortality (about 9% per year), while female mortality has increased slightly. Surgery, radiation therapy, and chemotherapy are Treatment options for pancreatic cancer. These treatment options can prolong the survival of many patients and/or alleviate the symptoms of many patients, but for most patients, it is unlikely to be cured. There are significant needs for other treatment and diagnostic options for the disease. These options include the use of antibodies, vaccines and small molecules as a form of treatment. In addition, in all areas of cancer treatment and research, these forms are also needed for diagnosis and detection. Measuring, monitoring and facilitating research tools of prior art. Therapeutic utility of monoclonal antibodies (mAb) (G. K〇Mer and c Milstein,

Nature 256: 495_497 (1975)) is learning. Individual antibodies have now been approved as a treatment for transplantation, cancer, infectious diseases, cardiovascular diseases and inflammation. Different isotypes have different effector functions. These functional differences are reflected in different three-dimensional structures of various immunoglobulin isotypes (p Μ

Alzari et al., Annual Rev. Immunol., 6:555-580 (1988)). Since mice are susceptible to immunization and can recognize most of the human antigens as foreign antigens, mAbs for human targets with therapeutic potential are generally derived from murine animals. However, the murine 111 8 15 has an inherent defect as a human therapeutic. Since mAbs have a shorter circulating half-life in humans than human antibodies', so they need to be more frequently given. More importantly, the repeated administration of I42769.doc -10· 201021828 murine antibodies to the human immune system causes the human immune system. The following reaction is produced: the mouse protein is recognized as a foreign protein and a human anti-mouse antibody (HAMA) reaction is produced. This HAMA response can cause allergic reactions and rapid clearance of murine antibodies from the system, rendering the murine anti-therapeutic use useless. To avoid these effects, efforts have been made to form the human immune system in mice. It was originally envisaged to form a transgenic mouse capable of responding to an antigen with an antibody having a human sequence (see Bruggemann et al., proc

® Nat'l. Acad. Sci. USA 86:6709-6713 (1989)), but limited by the amount of DNA, which can be stably maintained by the available selection vector. The use of a yeast artificial chromosome (YAC) selection vector serves as a guide for the introduction of large germline fragments of the human Ig locus into mammals of the transgenic gene. Most of the human V, D, and J region genes that are present at the same interval in the human genome and human constant regions are essentially introduced into the machine using YAC. The transgenic mouse product is called XenoMouse®. Rats are also available from Abgenix, Inc. (Fremont CA). SUMMARY OF THE INVENTION The present invention provides an antibody, a binding fragment thereof, and a molecule engineered thereof, which bind to a polypeptide fragment of 58P1D12 protein and 58P1D12 protein. The invention encompasses polyclonal and monoclonal antibodies, murine and other mammalian antibodies, chimeric antibodies, humanized and fully human antibodies, and antibodies labeled with a detectable label or therapeutic agent. In certain embodiments, the limiting conditions are that the entire nucleic acid sequence of Figure 3 is not encoded and/or the entire amino acid sequence of Figure 2 is not prepared. In some embodiments, the entire nucleic acid sequence encoding Figure 3 and/or the preparation of 142769.doc 201021828, the entire amino acid sequence of Figure 2, employs the corresponding human unit dosage form. The present invention further provides a method of detecting the presence and state of 58 piD12 polynucleic acid and protein in a plurality of biological samples, and a method of identifying cells expressing 5 8p 丨 D 丨 2 . One embodiment of the invention provides a method of monitoring a 58P1D12 gene product in a tissue or hematology sample having or suspected of having some form of growth disorder, such as cancer. The invention further provides a plurality of immunogenic or therapeutic compositions for use in the treatment of cancers exhibiting 58P1D12, such as cancers of the tissues listed in Table I, and the ❹ strategy includes the aim of inhibiting the transcription, translation, processing or function of 58P1D12 Therapy and cancer vaccine. In one aspect, the invention provides a composition for treating a cancer of 58P1D12 in a human subject, and a method comprising the same, wherein the composition comprises a carrier suitable for human use and one or more inhibiting the production or function of 58P1D12 The human unit dose of the agent. The carrier is preferably a human specific carrier. In another aspect of the invention, the agent is part that is immunoreactive with the 58P1D12 protein. Non-limiting examples of such moieties include, but are not limited to, antibodies (such as single chain antibodies, monoclonal antibodies, multi-drug antibodies, humanized antibodies, chimeric antibodies, or human antibodies), functional equivalents thereof (regardless of Naturally occurring or synthetic) and combinations thereof. The antibodies can be conjugated to a diagnostic or therapeutic moiety. In another aspect, the agent is a small molecule as defined herein. [Embodiment] Summary of each chapter I) Definition 142769.doc •12- 201021828 II.) 58P1D12 polynucleotide II. A.) Use of 58P1D12 polynucleic acid II.A.1.) Monitoring genetic abnormality II.A .2·) Antisense Example II.A.3.) Primer and primer pair II.A.4.) Isolation of nucleic acid molecule encoding 58piD12 III.)

IV·) Π.Α.5·) Recombinant Nucleic Acid Molecule and Host-Vector System 58P1D12 Related Protein III_ A.) Protein with Primitive Example III. B·) Expression of 58P1D12 Related Protein III.C·) 58P1D12 Related Protein Modification III.D.) 5 8P1D12 related protein use 58P1D12 antibody V.) 58P1D12 cellular immune response VI. ) 58P1D12 transgenic animal VII.) Method 58P1D12 for debt testing VIII. ) Monitoring the status of 58P1D12 related genes and their products Method IX) Identification of the molecule interacting with 58P1D12 X) Therapeutic methods and compositions XA) Anticancer vaccine XB·) 58P1D12 XC) 58P1D12 as a target for antibody-based therapy as a target for cellular immune responses XCI minigene (minigene) vaccine XC2. Combination of CTL peptide and helper peptide 142769.doc -13- 201021828 X · C · 3. Combination of CTL and T cell sensitizer XC4. Contains pulse labeling by CTL and/or HTL peptide Dc vaccine composition XD) adoptive immunotherapy XE) administration of vaccine for therapeutic or prophylactic purposes XI_) diagnostic and prognostic aspects of 58P1D12 Example XII. Inhibition of 58P1D12 protein function ΧΙΙ·Α_) Intracellular Inhibition of 58P1D12 ΧΙΙ.Β·) Inhibition of 58P1D12 by recombinant protein ◎ XII.C.) Inhibition of 58P1D12 transcription or translation of XII.D.) Treatment strategy · General considerations XIII.) Identification, characterization and use of 58P1D12 modulator XIV. Therapeutic Uses of RNAi and Small Interfering RNA (siRNA) XV.) Sets/Products I.) Definitions: Unless otherwise defined, all the specific terms, notes, and other scientific techniques used herein are intended to have the skill of the present invention. The meaning of what is usually understood. In some instances, terms that have a commonly understood meaning are defined in this document for clarity and/or for alternate reference, and the inclusion of such definitions herein is not necessarily to be construed as indicating that it has the meaning commonly understood in the art. Substantial difference. Many of the techniques and procedures described or referenced herein are well known to those skilled in the art and are commonly employed using conventional methods, such as the widely used methods of molecular selection described in the following literature: Sambrook et al., Molecular Cloning: A Lab〇rat〇ry 142769.doc -14- 201021828

Manual 2nd Edition (1989) Cold Spring Harbor Laborat〇ry press. Cold Spring Harbor, N.Y. Unless otherwise stated, procedures involving the use of commercially available kits and reagents are generally performed according to the manufacturer's specified protocol and/or parameters, as appropriate. The terms "advanced cancer", "locally advanced cancer", "advanced disease" and "locally advanced disease" mean cancer that has spread through the tissue of the relevant tissue and is intended to be included in the American Urological Society (American Urology).

Association; AUA) Systemic C disease; C1 _C2 disease according to the Whitmore-Jewett 10 system; and T3-T4 and N+ disease according to the TNM (tumor, nodule, metastasis) system. In general, surgery is not recommended for patients with locally advanced disease, and such patients have substantially less favorable outcomes than patients with clinically localized (organ-limited) cancer. " Altering the natural glycosylation pattern" is herein intended to mean the deletion of one or more carbohydrate moieties present in the native sequence 58P1D12 (by removing the basic glycosylation site, or by using chemistry and/or Alternatively or in an enzymatic manner, the glycosylation is deleted, and/or one or more glycosylation sites not present in the native sequence 58P1D12 are added. In addition, the phrase includes altering the glycosylation properties of the native protein, including altering the nature and proportion of the various carbohydrate moieties present. The term "analog" refers to a molecule that is structurally similar or shares similar or identical properties to another molecule (eg, 58p 1 d 12 related protein). For example, an analog of 5,58P1D12 protein can be specifically bound by an antibody or tau cell that specifically binds to 58P1D12. The term "antibody" is used in its broadest sense unless explicitly stated otherwise. 142769.doc 201021828 The second "antibody" may be a naturally occurring antibody or an artificial antibody, such as a monoclonal antibody prepared by conventional fusion tumor technology. anti-58piDi2 resistance

The body comprises a monoclonal antibody and a plurality of antibodies' and fragments of the antibody comprising an antigen binding domain and/or one or more complementarity determining regions. As used herein, the term "antibody" refers to any form of antibody or fragment thereof that specifically binds 58PID12 and/or exhibits the desired biological activity, and specifically encompasses monoclonal antibodies (including full-length monoclonal antibodies), multiple antibodies, Multispecific antibodies (eg, bispecific antibodies) and antibody fragments, as long as they specifically bind to 58 pim2 and/or exhibit the desired biological activity. Any specific antibody can be used in the methods and compositions provided herein. Thus, in one embodiment, the term "antibody" encompasses at least one molecule derived from the variable region of an immunoglobulin light chain molecule and at least one variable region derived from a heavy chain molecule, the variable regions being combined to form A specific binding site for the target antigen. In one embodiment, the antibody is an IgG antibody. For example, the antibody is an IgG1, Post 2, Post 3 or Post 4 antibody. Antibodies suitable for use in the methods and compositions of the invention can be produced in cell culture, phage, or various animals including, but not limited to, cows, rabbits, goats, mice, rats, hamsters, guinea pigs, sheep, dogs, Cats, monkeys, chimpanzees, baboons. Thus, in one embodiment, the antibody of the invention is a mammalian antibody. Phage technology can be used to isolate the original antibody or to produce variants with altered specificity or affinity properties. These techniques are common techniques and are well known in the art. In one embodiment, the anti-system is prepared by recombinant means known in the art. For example, a recombinant antibody can be produced by transfecting a host cell with a vector comprising a DNA sequence encoding the antibody. The DNA 142769.doc -16-201021828 sequence representing at least one VL and at least one VH region can be transfected into a host cell using one or more vectors. Examples of recombinant methods for antibody production and preparation

Demonstrative notes include Delves, ANTIBODY PRODUCTION: ESSENTIAL

TECHNIQUES (Wiley, 1997); Shephard et al., MONOCLONAL ANTIBODIES (Oxford University Press, 2000); Goding,

MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE

(Academic Press, 1993); CURRENT PROTOCOLS IN IMMUNOLOGY (John Wiley & Sons, latest version). The antibodies of the invention can be modified by recombinant means to enhance the greater efficacy of the antibody in mediating the desired function. Thus, it is within the scope of the invention to modify the anti-system by substitution using recombinant means. Substitutions are usually conservative substitutions. For example, at least one amino acid in the constant region of the antibody can be substituted with a different residue. See, e.g., U.S. Patent No. 5,624,821, U.S. Patent No. 6,194,551, Application No. WO 9958572, and Angal et al., Mol. Immunol. 30: 105-08 (1993). The modification of the amino acid includes the deletion, addition, and substitution of the amino acid. In some cases, such changes are intended to reduce undesired activity, such as complement dependent cytotoxicity. Antibodies are typically labeled by covalent or non-covalent attachment to a substance that provides a detectable signal. A wide variety of marking and joining techniques are known and are widely reported in scientific and patent literature. Screening of these antibodies in combination with normal 58P1D12 or defective 58P1D12 can be performed. See, for example, ANTIBODY ENGINEERING: A PRACTICAL APPROACH (Oxford University Press, 1996). Appropriate antibodies having the desired biological activity may be identified by in vitro assays including, but not limited to, the following assays: proliferation, migration, adhesion, soft agar growth, angiogenesis, cell-cell communication, apoptosis, transfection 142769.doc • 17- 201021828 Transmitted with h, and can be identified by the following in vivo assays, such as tumor growth inhibition. The antibodies provided herein can also be used in diagnostic applications. As a capture antibody or a non-neutralizing antibody, the ability to bind to a specific antigen without inhibiting the receptor binding or biological activity of the antigen can be screened. As neutralizing antibodies, such antibodies can be used in competitive binding assays. It can also be used to define #58P1D12 or its receptor. An antibody fragment" is defined as at least a portion, i.e., an antigen binding region, of a variable region of an immunoglobulin molecule that binds to its target. In the examples, it specifically covers a single anti-58P1D12 antibody and its pure lines (including agonists, antagonists, and neutralizing antibodies) and anti-12 steroid compositions having multiple epitope specificity. The antibodies of the methods and compositions of the invention may be monoclonal or polyclonal antibodies. The antibody may comprise a Fab fragment, an F(ab')2 fragment, a single chain variable region, and the like in the form of an antibody fragment that binds to the antigen. The tablets of the Jade Knife can be produced using methods well known in the art and include enzymatic decomposition and recombination. As used herein, any form of "antigen" can be utilized to produce antibodies specific for 58PID12. Thus, the elicited antigen can be a single anti-prostaglandin, a multiplex epitope, or a complete protein alone or in combination with one or more immunogenic enhancers known in the art. The elicited antigen can be a full-length protein, a cell surface protein (eg, only "immunized with at least a portion of the star antigen) or a soluble protein (eg, only the extracellular domain of the protein is used for immunization) ). The antigen can be prepared using genetically engineered cells. The dna encoding the antigen may be genomic DNA or non-genomic DNA (e.g., cDNA) and encode at least the extracellular domain I42769.doc • 18· 201021828 - part. The term "portion" as used herein refers to the minimum number of amino acids or nucleic acids that, if appropriate, constitute the immunogenic epitope of the desired antigen. Any deportable vector suitable for the cells required for transformation can be used, including, but not limited to, adenoviral vectors, plastids, and non-viral vectors such as yang = daughter lipids. In the examples, the antibodies of the methods and compositions herein specifically bind at least a portion of the extracellular domain of 58P1D12 required.

The antibody or antigen-binding fragment thereof provided herein can be conjugated to a "bioactive agent". As used herein, the term "biologically active agent" refers to any synthetic compound or naturally occurring compound that binds to an antigen and/or enhances or mediates a desired biological effect to enhance the toxin that kills the cell. In one embodiment, a binding fragment suitable for use in the present invention is a biologically active fragment. As used herein, the term "biologically active" means that the antibody or antibody fragment is capable of binding to the epitope of the desired antigen and exerts a biological effect, either directly or indirectly. Direct effects include, but are not limited to, modulation of 'stimulation and/or inhibition of growth signals; regulation, stimulation, and/or inhibition of anti-apoptotic signals; regulation, stimulation, and/or inhibition of apoptosis or necrosis signals; regulation, stimulation, and / or inhibit the ADCC cascade; and regulate, stimulate and/or inhibit the CDC cascade. "Bispecific" antibodies are also suitable for use in the methods and compositions of the invention. As used herein, the term "bispecific antibody" refers to an antibody having binding specificity for an antigenic thiol group of at least two different antigens, usually a monoclonal antibody. In one embodiment, the epitope is derived from the same antigen. In another embodiment the ' epitope is derived from two different antigens. Methods for making bispecific antibodies are known in the art. For example, a bispecific antibody can be recombinantly produced using the co-expression of two pairs of immunoglobulin heavy/light chains. See 142769.doc • 19· 201021828 eg Milstein et al., Nature 305:537-39 (1983). Alternatively, bispecific antibodies can be prepared using chemical linkages. See, for example, Brennan et al., Science 229: 81 (1985). Bispecific antibodies include bispecific antibody fragments. See, for example, Hollinger et al, proc. Natl. Acad. Sci. USA 90:6444-48 (1993), Gruber et al, j. Immun〇1 152:5368 (1994)° The monoclonal antibodies herein include, inter alia, r-embedded An antibody wherein one of the heavy and/or light chain is identical or homologous to a corresponding sequence of an antibody derived from a particular species or belonging to a particular antibody or subclass, and the remainder of the (and other) strand is derived from another The corresponding sequences of a species or antibodies belonging to another antibody or subclass are identical or homologous; and fragments of such antibodies, as long as they specifically bind to the target antigen and/or exhibit the desired biological activity (U.S. Patent No. 4,816,567; And Morrison et al., Pr〇c Natl Acad % USA 81: 6851-6855 (1984)). The term "chemotherapeutic agent" refers to all compounds which are effective in inhibiting tumor growth. Non-limiting examples of chemotherapeutic agents include: alkylating agents such as nitrogen nitrene, ethyleneimine compounds and alkyl sulfonates; antimetabolites such as folic acid, guanidine or pyrimidine antagonists; mitotic inhibitors such as periwinkle Bioassay and podophyllotoxin derivatives; cytotoxic antibiotics; compounds that disrupt or interfere with DNA expression; and growth factor receptor antagonists. In addition, chemotherapeutic agents include cytotoxic agents (as defined herein), antibodies, biomolecules, and small molecules. The term "codon-optimized sequence" refers to a nucleotide sequence optimized for a particular host species, by replacing any codon having a frequency of use of less than about 20%. 142769.doc -20-201021828. For the optimization of a given host species, the elimination of 彳g平晚## See the optimization of the nuclear (four) sequence in the code _ ” 除 转 转 转 转 转 / / / 优化 优化 优化 优化 优化 优化 优化 优化 优化 优化Performance enhancement sequence." ,

"Lite library" is a collection of various compounds produced by a variety of chemical "basic components" (such as test H, chemical synthesis or biosynthesis. For example, linear combinatorial chemical libraries (such as peptides (such as For example, a mutant protein) library is formed for each given length of the compound (ie, the number of amino acids in the polypeptide compound) by combining a group of chemically basic components called amino acids in every possible manner. This combination of chemical basic components is combined to synthesize a variety of compounds (Gauop et al, J Med Chem 37 (9): 1233-1251 (1994).) Preparation and screening of combinatorial libraries are well known to those skilled in the art. Such combinatorial chemical libraries include, but are not limited to, peptide libraries (see, e.g., U.S. Patent No. 5,010,175; Furka, Pept. Prot. Res. 37:487-493 (1991) Houghton Specialist' Nature, 3 54:84-88 (1991)), genus (pct publication No. W〇91/19735), coding peptide (PCT Publication WO 93/20242), random biological oligomer (PCT Publication WO 92/00091), benzo Benzodizepine (US Patent No. No. 5,288,514), diversomer (such as beta-urea, benzodiazepine and dipeptide) (H〇bbs et al, Proc. Nat. Acad. Sci. USA 90: 6909-6913 (1993)) , vinylogous polypeptide (Hagihara et al., J. Amer.

Chem. Soc. 114:6568 (1992)), a non-peptide peptidomimetic having a β-D-glucose backbone (Hirschmann et al., J. Amer. Chem. Soc. 114: 9217-142769.doc -21 - 201021828 9218 (1992)), similar organic synthesis of small compound libraries (Chen et al. 'J. Amer. Chem. Soc. 116:2661 (1994)), oligoamino phthalate (Cho et al., Science 261:1303 ( 1993)) and / or peptidyl phosphonates (Campbell et al, J. 〇rg. Chem. 59: 658 (1994)). See, in general, Gordon et al, J. Med. Chem. 37: 1385 (1994); nucleic acid libraries (see, for example, Stratagene, Corp.), peptide nucleic acid libraries (see, e.g., U.S. Patent 5,539,083), antibody libraries (see, e.g., Vaughn et al. Nature Biotechnology 14(3): 309-314 (1996), and PCT/US96/10287), carbohydrate libraries (see, for example, Liang et al. '8 (^11〇6 274:1520-1522 (1996), and US patents) No. 5,593,853) and a library of small organic molecules (see, for example, benzodiazepine: Baum, C & EN, Jan. 18, p. 33 (1993); isoprenoid: U.S. Patent No. 5,569,588; Thiazolidinone and metathiazanone: U.S. Patent No. 5,549,974; pyrrolidine: U.S. Patent Nos. 5,525,735 and 5,519,134; morpholinyl compound: U.S. Patent No. 5,506,337; Diazol: U.S. Patent No. 5,288,514; and its analogs. Devices for preparing combinatorial libraries are commercially available (see, for example, 357 NIPS, 390 NIPS, Advanced Chem Tech, Louisville KY; Symphony, Rainin, Woburn, ΜΑ; 433A, Applied Biosystems, Foste r City, CA; 9050, Plus, Millipore, Bedford, NIA. A variety of well-known robotic systems for solution phase chemistry have also been developed. These systems include automated workstations such as Takeda Chemical Industries, LTD. (Osaka, Japan). The automated synthesis equipment developed and the robotic system using the robotic arm to simulate the synthetic operations performed by the pharmacist 142769.doc -22- 201021828 (Zymate H, Zymark Corporation, Hopkinton, Mass.; Orca, Hewlett-Packard, Palo Alto Any of the above devices are suitable for use in the present invention. The nature and improved implementation of such devices, if any, so that they can operate as described herein, will be apparent to those skilled in the art. A variety of combinatorial libraries are commercially available (see, for example, ComGenex, Princeton, NJ; Asinex, Moscow, RU; Tripos, Inc., St. Louis, MO; ChemStar, Ltd, Moscow, RU; 3D Pharmaceuticals, Exton, PA; Martek Biosciences , Columbia, Lu MD, etc.). As used herein, the term "conservative substitution" means that the amino acid substitution is known to those skilled in the art and can generally be carried out without causing a change in the biological activity of the resulting molecule. Those skilled in the art will appreciate that single amino acid substitutions in non-essential regions of the polypeptide will generally not substantially alter biological activity (see, for example, Watson et al., MOLECULAR BIOLOGY OF THE GENE, The Benjamin/Cummings Pub. Co., p. 224 (4th edition, 1987)). These exemplary substitutions are preferably carried out according to the substitutions listed in Table 111 (a-b). For example, the alterations include the substitution of any of the other hydrophobic amino acids with any one of isoleucine (I), valine (V), and leucine (L); Substituting aspartic acid (D) for glutamic acid (E) and vice versa; replacing glutamine (N) with glutamine (Q) and vice versa; and replacing with serine (S) Threonine (T) and vice versa. Depending on the environment of the particular amino acid and its role in the three-dimensional structure of the protein, other substitutions may also be considered conservative substitutions. For example, glycine (G) and alanine (A) are usually interchangeable, while alanine (A) and valine (V) are also interchangeable. 142769.doc -23-201021828. The thiamine (M) which is relatively hydrophobic is generally interchangeable with leucine and isoleucine, and sometimes it is interchangeable with niacin. The lysine (8) and arginine (R) are generally interchangeable in positions where the significant character of the amino acid residue is its charge and the pK of the two amino acid residues are not important. Other changes can be considered “conservative” in special circumstances (see, for example, Table 111(a) in this article; “Biochemistry”, second edition, page 13_15, [Xian Wei

Stryer (Stanford University); Henikoff et al, PNAS 1992, Vol. 89, 10915-10919; Lei et al, j Biolchem, May 19, 1995; 270(20): 11882-6). Other substitutions are also permissible and may be judged on the basis of a test or in accordance with known conservative substitutions. The term "cytotoxic agent" refers to a substance that inhibits or prevents the expression of cells, the function of cells, and/or the destruction of cells. This term is intended to include radioisotopes, chemotherapeutics, and toxins, such as small molecule toxins or enzymatically active toxins, including fragments and/or variants thereof, of bacterial, fungal, plant or animal origin. Examples of cytotoxic agents include, but are not limited to, Orlista> Auristatin, Orlista, Auristatin e, auromycin, maytansinoid, 钇, silver, ricin G, ricin A, cobrastatin, duocarmycin, dolostatin, doxorubicin, Daunorubicin, taxol, cisplatin, cc 1065, etidium bromide, mitomycin, etoposide, teno Tenoposide, vincristine, vinblastine, colchicine, anthracnose II 142769.doc •24- 201021828 _ (dihydroxy anthracin dione), actinomycin (actinomycin), diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin (abrin), abrin toxin A chain, modeccin A chain , alpha-sarcin, gelonin, have Mitigellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin ), Sapaonaria officinalis inhibitors and glucocorticoids and other chemotherapeutic agents, and radioisotopes such as At211, I131, I125, Y90, Re丨86, Re188, Sm (5), Bi2丨2 or Bi2丨3, Radioisotopes of p3 and Lu, including Lu177. The antibody may also be conjugated to an anti-cancer prodrug activating enzyme capable of converting the prodrug into its active form. As used herein, the term "bifunctional antibody" refers to a small antibody fragment having two antigen binding sites comprising a heavy chain variable domain (Vh) linked by the same polypeptide chain (Vh-Vl). With the light chain variable domain (Vl). By using a linker that is too short to allow pairing between the two domains on the same strand, the domains are forced to pair with the complementary domains of the other strand and form two antigen-binding sites. Bifunctional antibodies are more fully described in, for example, Ep 404,097; WO 93/11161; and Hollinger et al, Proc. Natl Acad. Sci. USA 90:6444-48 (1993). "Gene product" is used herein to mean peptide/protein* mRNA. For example, "the gene product of the present invention" is sometimes referred to herein as "cancer amino acid sequence", "cancer protein", and the protein of cancer listed in the table 142769.doc -25- 201021828" "Cancer mRNA", "mRNA of cancer listed in the table", etc. In the implementation of the 'Fruit' cancer protein line is encoded by the nucleic acid of Figure i. The cancer protein can be a fragment or a full length protein encoded by the nucleic acid of the map. In an embodiment, the cancer amino acid sequence is used to determine sequence- or similarity. In another implementation, the sequences are naturally occurring dual gene variants of the protein encoded by the nucleic acid of Figure k. In another embodiment, the sequences are sequence variants as further described herein. Heterotypic antibodies can be used in the methods and compositions of the invention. As used herein, the term "heterotypic antibody" refers to two covalently linked antibodies. Such antibodies can be prepared by methods known in the art of synthetic protein chemistry, including the use of cross-linking agents. See, for example, U.S. Patent No.. High throughput screening assays for the presence, absence, quantification or other properties of a particular nucleic acid or protein product are well known to those skilled in the art. Similarly, binding assays and reporter gene assays are similarly well known. Thus, for example, U.S. Patent No. 5,559,410 discloses high-throughput screening for proteins; U.S. Patent No. 5,585,639 discloses high-throughput screening for nucleic acid binding (i.e., arrays); and U.S. Patent No. 5,576,220 And No. 5,541,061 discloses a high throughput assay for ligand/antibody binding. In addition, high-throughput screening systems are commercially available (see, for example, Amersham Biosciences, Piscataway, NJ; Zymark Corp., Hopkinton, MA; Air Technical Industries, Mentor, OH; Beckman Instruments, Inc. Fullerton, CA; Precision Systems, Inc. ., Natick, MA, etc.). These systems typically automate all procedures, including 142769.doc -26- 201021828 with sample and reagent pipetting, liquid dispensing, timing incubation, and final reading of the microplate with a detector suitable for assay. These configurable systems offer high throughput and fast start-up as well as a high degree of flexibility and customization. Manufacturers of these systems offer detailed solutions for a variety of high-throughput systems. Thus, for example, Zymark Corp. provides a technical bulletin describing a screening system for the regulation of debt detection, ligand binding, and the like. The term "homolog" refers to a molecule that exhibits homology to another molecule, for example, a sequence of chemical residues that are identical or similar at the corresponding positions. In one embodiment, the antibody provided herein is a "human antibody." As used herein, the term "human antibody" refers to an antibody in which the entire sequence of the light and heavy chain sequences, including the complementarity determining regions (CDRs), is substantially derived from a human gene. In one embodiment, a human monoclonal antibody system is produced by the following techniques: trioma technology, human B cell technology (see, eg, Kozbor et al, Immunol. Today 4: 72 (1983)), EBV transformation technology (See, for example, Cole et al. MONOCLONAL ANTIBODIES AND CANCER THERAPY 77-96 (1985)) or presented using sputum bacteria (see, eg, Marks et al, J. Mol. Biol. 222: 581 (1991)). In a specific embodiment, a human antibody is produced in a transgenic mouse. Techniques for preparing such portions to fully human antibodies are known in the art and any such techniques can be used. According to a particularly preferred embodiment, a fully human antibody sequence is produced in a transgenic mouse engineered to express human heavy and light chain antibody genes. An exemplary description of the preparation of a transgenic mouse that produces a human antibody can be found in application No. WO 02/43478 and U.S. Patent 6,657,103 (Abgenix) and its succession. B cells which produce a transgenic mouse which is intended to be resistant to 142769.doc -27-201021828 can then be fused to prepare a fusion tumor cell line for continuous production of antibodies. See, for example, U.S. Patent Nos. 5,569,825; 5,625,126; 5,633,425; 5,661,016; and 5,545,8063⁄4^.; AJakobovits, Adv. Drug Del. Rev. 31:33-42 (1998); J. Exp. Med. 188: 483-95 (1998) ° "Human leukocyte antigen" or "HLA" is a human class I or class II major histocompatibility complex (MHC) protein (see, for example, Stites et al. IMMUNOLOGY, 8th ed., Lange Publishing, Los Altos, CA (1994)). As used herein, the term "humanized antibody" refers to an antibody form comprising sequences from non-human (e.g., murine) antibodies as well as human antibodies. Such antibodies are chimeric antibodies which contain minimal sequences derived from non-human immunoglobulins. Generally, a humanized antibody comprises substantially all of at least one and usually two variable domains, wherein the hypervariable loops all or substantially all correspond to the sequence of the non-human immunoglobulin and the FR regions are all or substantially All are their sequences of human immunoglobulin sequences. The humanized antibody also includes at least a portion of an immunoglobulin (typically a human immunoglobulin) constant region (Fc), as desired. See, for example, Cabilly, U.S. Patent No. 4,816,567; Queen et al. (1989) Proc. Nat'l Acad. Sci. USA 86:10029-10033; and ANTIBODY ENGINEERING: A PRACTICAL APPROACH (Oxford University Press 1996) o in nucleosides The term "hybridization" and the like in the context of acid are intended to refer to conventional hybridization conditions, preferably such as hybridization in 50% guanamine/6XSSC/0.1% SDS/100 pg/ml ssDNA, wherein the hybridization temperature exceeds 142769.doc • 28 · 201021828 The temperature at 37 ° C and washed in 0.1XSSC/0.1% SDS exceeds 55〇c. The word "isolated" or "biologically pure" means that the substance is substantially or substantially free of components that normally accompany the substance when it is in its natural state. Thus, the isolated peptide of the present invention preferably does not contain materials which are normally associated with such identities in its original environment. For example, when the polynucleic acid is substantially separated from a contaminating polynucleotide corresponding to or complementary to a gene other than the 58P1D12 gene or a polypeptide encoding a polypeptide other than the 58P1D12 gene product or a fragment thereof, Glycosylates are "isolated." Those skilled in the art can readily obtain isolated 58P1D12 polynucleic acid using a nucleic acid isolation procedure. For example, a protein is said to be "isolated" when it is physically, mechanically or chemically removed from the cellular components normally associated with the protein. Those skilled in the art can readily obtain the isolated 58P1D12 protein using standard purification methods. Alternatively, the isolated protein can be prepared chemically. Suitable "markers" include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent moieties, chemiluminescent moieties, magnetic particles and the like. Patents that teach the use of such indicia include U.S. Patent Nos. 3,817,8,7, 3,850,752, 3,939,350, 3,996,345, 4,277,437, 4,275,149, and 4,366,241. Furthermore, the antibodies provided herein can be used as an antigen binding component of the phosphor (1) uorob〇dy). See, for example, Zeytun et al., Nat. Biotechnol. 21:1473-79 (2003). The term "mammal" refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses, and humans. In one embodiment of the invention 142769.doc • 29-201021828, the mammal is a mouse. In another embodiment of the invention, the mammal is a human. The terms "metastatic cancer" and "metastatic disease" mean a cancer that has spread to a regional lymph node or has spread to a distal site, and is intended to include a stage D disease according to the AUA system and a ΤχΝχΜ+ stage disease according to the sputum system. The term "modulator" or "test compound" or "candidate drug" or grammatically equivalent terms as used herein is intended to directly or indirectly alter the expression of a cancer phenotype or cancer sequence (eg, a nucleic acid or protein sequence) or Any molecule that has the ability to test for cancer sequence effects (eg, signal transduction, gene expression, protein interaction, etc.), such as proteins, oligopeptides, small organic molecules, polysaccharides, polynucleotides, and the like. In one aspect, the modulator neutralizes the effects of the cancer protein of the invention. "Neutralization" means inhibiting or blocking the activity of the protein and its subsequent effects on the cell. In another aspect, the modulator neutralizes the effect of the gene of the invention and neutralizes the effect of the protein by normalizing the content of the corresponding protein. In a preferred embodiment, the modulator alters the performance profile, or the performance profile of the nucleic acid or protein provided herein, or the downstream effector pathway. In an embodiment, the modulator inhibits cancer phenotype to, for example, a normal tissue fingerprint. In another embodiment, the modulator induces a cancer phenotype. A plurality of assay mixtures are typically tested in parallel at different drug concentrations to obtain differential responses for various waves. These concentrations are usually used as negative controls, ie below the zero concentration or detection limit. Modulators, drug candidates or test compounds encompass many chemical classes but are typically organic molecules 'preferably small organic compounds having a molecular weight greater than (10) Daltons (eg, 142769.doc -30.201021828 and less than about 2,500 Daltons). Preferably, the small molecule is less than 2000 D or less than 1500 D or less than 1 〇〇〇 D or less than 5 〇〇 d. The candidate agent comprises functional groups necessary for structural interaction (especially hydrogen bonding) with the protein, and usually includes at least Or an amine group, a benzyl group, a thio group or a hydryl group, preferably at least two of the chemical functional groups. The candidate agent typically comprises a ring carbon or heterocyclic ring structure substituted with one or more of the above functional groups and/or Aromatic or polyaromatic structure. Modulators also contain biomolecules such as peptides, sugars, fatty acids, steroids, sputum, shouting, and h 4仏.,,., ^ Preferably, the peptide is particularly preferred. The class-modulating agent is, for example, having from about 5 to about 35 amino acids, preferably from about 5 to about 20 amino acids, and particularly preferably from about 7 to about 15 amino acids. Peptide. Cancer regulatory protein is preferably soluble, including Non-transmembrane regions and/or have a soluble terminal Cys. In one embodiment, the C-terminus of the fragment is maintained as a free acid and the end is conjugated (ie, coupled to cysteine) a free amine. In one embodiment, the cancer protein of the invention is conjugated to an immunogenic agent as discussed herein. In & one, the cancer protein is conjugated to BSA. The peptide of the invention (e.g., preferably The lengths can be linked to each other or to other amino acids to form longer peptides/proteins. The regulatory peptide can be a digest, a random peptide or a "heavier" random peptide of a naturally occurring protein as described above. In a preferred embodiment, the peptide/protein based modulator is an antibody and fragment thereof as defined herein. The cancer modulator may also be a nucleic acid. The nucleic acid modulator may be a naturally occurring nucleic acid, a random nucleic acid or a "heavy" random nucleic acid. For example, the digest of the prokaryotic or eukaryotic genome can be used analogously to the method described above for the method of protein 142769.doc • 31 · 201021828. As used herein, the term “ "Strain" refers to an antibody obtained from a population of substantially the same antibody 'that is, in addition to a small number of possible naturally occurring mutations, 'the individual antibodies that make up the population are identical. The monoclonal antibody has an epitope against a single antigen. Highly specific. In contrast, conventional (multiple) antibody preparations typically include multiple antibodies directed against (or specific for) different epitopes. In one embodiment, multiple antibodies contain multiple monoclonal antibodies' The monoclonal antibodies have different epitope specificity, affinity or affinity in a single lactogen containing multiple antigenic epitopes. The modifier "single plant" means obtained from substantially the same antibody sound. The nature of antibody 1 and should not be construed as requiring preparation of the antibody by any particular method. For example, a monoclonal antibody to be used in accordance with the present invention can be prepared by the fusion tumor method first described by Kohler et al, Nature 256: 495 (1975), or by recombinant DNA methods (see, for example, US patents) Preparation No. 4, 8 16, 567). "Single antibody" can also be isolated from a phage antibody library using, for example, the techniques described in Clacks 〇n et al.

Nature 352: 624-628 (1991) and Marks et al, j. Mol. Bi〇1 222: 581-597 (1991). Such monoclonal antibodies are typically at least about 1 μΜ, more typically at least about 3 μM, usually at least about 30 nM, preferably at least about 1 μM, more preferably at least about 3 nM, or more, as determined by ELISA. Good Kd combination. "primitive" (eg, in the biological basis of the 58P1D12-related protein) refers to any amino acid form that forms part of the sequence: interaction with a specific function (eg, protein-protein interaction, protein f_DNA interaction, etc.) or repair 142769.doc •32- 201021828 A sequence of proteins associated with (eg, phosphorylation, glycosylation, or guanidine) or localization (eg, secretory sequences, nuclear localization sequences, etc.), or with the presence of immunogenicity (corporeal fluid or cells) ) related sequences. Primitives may be B-connected or capable of being aligned with certain locations. These locations are typically associated with a certain function or characteristic. In the context of an HLa motif, "primitive" refers to a peptide of a specified length that is recognized by a particular HLA molecule (typically a peptide having from about 8 to about 13 amino acids for a class I HLA motif; And for the η-type hl A motif, it is typically a residue pattern in a peptide having from about 6 to about 25 amino acids. For various proteins encoded by various human ® hla alleles, the peptide motifs for HLA binding are generally different, and the patterns of primary anchor residues and secondary anchor residues vary. The frequently occurring primitives are listed in Table V. "Pharmaceutical excipients" include substances such as adjuvants, carriers, pH adjusters and buffers, tonicity regulators, wetting agents, preservatives and the like. "Pharmaceutically acceptable" means a composition that is non-toxic, inert, and/or physiologically compatible with humans or other mammals. The term "polynucleotide" means a polymeric form of a nucleotide of at least 1 base or base pair (a modified form of a ribonucleotide or a deoxynucleotide or a nucleotide of either type). And is intended to include both single-stranded and double-stranded forms of DNA and/or RNA. In the art, the term is generally used interchangeably with "oligonucleotide J. Polynucleotides may comprise the nucleotide sequences disclosed herein, wherein, for example, thymidine (D) as shown in Figure 1 may also be used. Is uracil (u); this definition is related to the difference between the chemical structure of DNA and RNA, especially related to the following observations. · One of the four major bases in RNA is uracil (cut 142769.doc • 33· 201021828 Non-thymidine (τ). The term "polypeptide" means a polymer having at least about 4, 5, 6, 7 or 8 amino acids. Throughout the specification, the standard three-letter of amino acid is used. Or the name of the single parent. In this technique, the term is usually used interchangeably with "peptide" or "protein." HLA-ordered 疋 residue is an amino acid located at a specific position along the peptide sequence, which should be understood. To provide a point of contact between the immunogenic peptide and the HLA molecule. For immunogenic peptides, one to three (usually two) primary anchor residues define a "primary element" within a peptide of a specified length. Residues are understood to be in close contact with the peptide-binding groove of the HLA molecule. The side chain is embedded in a specific pocket of the binding groove. In one embodiment, for example, a class 1 anchoring residue of a class 1 hla molecule is located at a peptide antigen of 8, 9, 1 , η or 12 residues of the invention. The position of the base 2 (relative to the position of the terminal end of the amine) and the position of the terminal of the carboxy group are determined. Alternatively, in another embodiment, the tin sulfonium residue of the peptide which binds to the steroidal HLA molecule is spaced apart from each other, rather than the end of the peptide. Wherein the peptide is generally at least 9 amino acids in length. One of the motifs and one of the supermotifs is listed in Table IV(a). For example, a similar peptide can be altered by changing the values in Table IV. The presence or absence of a particular residue in the primary and/or secondary anchor sites is used to modulate the binding affinity and/or population coverage of a peptide comprising a particular HLA motif or supermotif. "Radioisotopes" include, but are not limited to, the following radioisotopes (non-limiting exemplary uses are also listed in Table IV (I)). As used herein, "randomization" or grammatical equivalence for nucleic acids and proteins. I have a substantially random nucleus Acids and Amines 142769.doc 34· 201021828 Acid Composition. These random peptides (or nucleic acids, as discussed herein) can be in any position and have any nucleotide or amino acid. Synthetic methods can be designed to generate randomization Protein or nucleic acid to allow formation of all or most of the possible combinations of all sequence lengths to form a library of randomized candidate protein bioactive agents. In the examples, the library is "completely randomized" and does not exist at any position The sequence is preferred or constant. In another embodiment, the library is a "frequently random" library. That is, some positions within the sequence remain constant, or are selected from a limited number of possibilities. For example, nucleotides or amine groups Acid residues are randomized within a defined class, such as hydrophobic amino acids, hydrophilic residues, spatially biased (small or large) residues, biased to form nucleic acid binding domains, form cysteine (for paternal associations), Amine acid (for SH-3 domain), serine acid, threonine, glutamic acid or histidine (for sulphation sites), etc., or formation of hydrazine or the like. A "recombinant" DNA or RNA molecule is a DNA or RNA molecule that has been manipulated by an in vitro molecule. As used herein, the term "single-chain Fv" or "r scFv" or "single-chain" anti-system refers to antibody fragments comprising the VH domain and the VL domain of an antibody, wherein such regions are present in a single polypeptide chain. The Fv polypeptide typically further comprises a polypeptide linker between the VH domain and the VL domain which enables the sFv to form the desired structure for antigen binding. For a review of sFv, see Pluckthun, THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, 113, ed., Rosenburg and Moore, Springer-Verlag, New York, pp. 269-315 (1994). Non-limiting examples of "small molecules" include compounds that bind to 58P1D12 or interact with 142769.doc-35-201021828 58P1D12, ligands that bind to and preferably inhibit 581>1〇12 protein functions, including hormones, neuropeptides, and tendencies. Chemical factors, odorants, linoleum and their functional equivalents. Preferably, the non-limiting small molecules have a molecular weight of less than about 10 kDa, more preferably less than about 9 kDa, about 8 kDa, about 7 kDa, about 6 kDa, about 5 kDa, or about 4 kDa. In certain embodiments, small molecules that are physically associated with or bind to the 58P1D12 protein are not found in naturally occurring metabolic pathways; and/or are more soluble in aqueous solutions than in non-aqueous solutions. As used herein, the term "specificity" refers to an antibody that selectively binds to an antigenic epitope of a target antigen. The binding specificity of the s-type antibody is determined by comparing the binding of the antibody to a suitable antigen under a given set of conditions and binding to an unrelated antigen or mixture of antigens. A binding of an antibody to an appropriate antigen is considered to be specific if it is at least 2-fold, 5-fold, 7-fold, and preferably 10-fold greater than the binding of the antibody to an unrelated antigen or mixture of antigens. In one embodiment, the specific antibody is an antibody that binds only to the 58P1D12 antigen but does not bind to an unrelated antigen. In another embodiment, the specific antibody is 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94% bound to the human 58P1D12 antigen but not bound to the 58P1D12 antigen. 95%, 96%, 97%, 9 8°/. , 99°/. Or an antibody to a non-human 58P1D12 antigen having 99% or more amino acid homology. In another embodiment, the specific antibody is an antibody that binds to the human 58P1D12 antigen and binds to the murine 58P1D12 antigen but has a higher degree of binding to the human antigen. In another embodiment, the specific antibody is an antibody that binds to the human 58P1D12 antigen and binds to the primate 58P1D12 antigen but has a higher degree of binding to the human antigen. In another embodiment, 142769.doc-36-201021828 and any non-human binding degree, the specific antibody binds to the human 58P1D12 antigen 58P1D12 antigen, but the "stringency" of the hybridization reaction with the human antigen or any combination thereof is easy. It is generally determined by the average technician and is empirically calculated based on the length of the probe, the temperature of the wash, and the salt concentration. In general, the longer the probe, the higher the temperature required for proper bonding, and the shorter the probe, the lower the temperature required. When the complementary strands are present below their refining temperature, the rhododendron reaction is generally dependent on the ability of the denatured nucleic acid sequence to re-adhere. The higher the desired degree of homology between the probe and the hybridizable sequence, the higher the relative temperature that can be used. Therefore, higher relative temperatures tend to make the reaction conditions more stringent, while lower temperatures do not. For additional details and explanations regarding the stringency of hybridization reactions, see Ausubel et al, Current Pr〇t〇c〇ls in Molecular Biology, Wiley Interscience Publishers, (1995). "Stringent conditions" or "high stringency conditions" as defined herein are as follows (but not limited to) identification: (1) use of low ionic strength and high temperature during washing, for example, using 0.015 Μ chlorination at 50 ° C Sodium / 0.0015 Μ sodium citrate / 0.1% sodium lauryl sulfate; (2) a denaturant (such as formamide, such as 50% (v / v) formamide) at 42 ° C during hybridization With 〇.1〇/0 bovine serum albumin / 0.1 ° /. Ficoll/〇.l% polyvinylpyrrolidone/5〇mM sodium phosphate buffer (pH 6.5) and 750 mM sodium hydride, 75 mM sodium citrate; or (3) 50% at 42 °C Indoleamine, 5xSSC (0.75 M NaCl, 0.075 citrate sodium citrate), 50 mM sodium sulphate (pH 6.8), 0.1% sodium pyrophosphate, 5 DDenhardt's solution, supersonic treatment鲑鱼精142769.doc -37· 201021828 DNA (50 pg/ml), 0.1% SDS and 10% dextran sulfate, washed at 0.2 ° SSC (sodium chloride / sodium citrate) at 42 ° C and Washing was carried out in 50% formamide at 55 ° C, followed by high stringency washes consisting of O.lxSSC containing EDTA at 55 °C. "Medium stringent conditions" are described, but are not limited to, in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and include washing solutions and hybridization conditions using stringent tb conditions as described above. (eg temperature, ionic strength and % SDS). An example of moderately stringent conditions is overnight incubation at 65 ° C in a solution containing the following: 1% bovine serum albumin, 0.5 M# sodium (pH 7.5), 1·25 mM EDTA, and 7% SDS/ 5xSSC (150 m M NaCl, 15 mM trisodium citrate), followed by washing of the filter in 2xSSC/l% SDS at 50 °C and washing the filter in 0.2x SSC/0.1% SDS at 50 °C. Those skilled in the art should understand how to adjust temperature, ionic strength, etc. as needed to accommodate factors such as probe length and similar factors. HL A "supermotif" is a peptide binding specificity shared by HLA molecules encoded by two or more HL A alleles. The overall phenotypic phenotypes of HLA supertypes for different populations are listed in Table IV (f). The non-limiting components of the various supertypes are as follows: A2: A*0201, A*0202, A*0203, A*0204, A*0205, A*0206 'A*6802, A*6901, A*0207 A3 : A3 , All , A31 , A*3301 , A*6801 , A*0301 , A*1101 , A*3101 B7 : B7 , B*3 501-03 , B*51 , B*5301 , B*5401 , 142769.doc -38- 201021828 B*5501 , B*5502 , B*5601 , B*6701 , B*7801 , B*0702 , B*5101 ' 6^5602 B44 : B*3701 , B*4402 , B* 4403, B*60 (B*4001), B61 (B*4006) A1 : A*〇l〇2, A*2604, A*3601, A*4301, A*8001 A24 : A*24, A*30 , A*2403, A*2404, A*3002, A*3003 B27: B*1401-02, B* 1503, B*1509, B*1510, ® B*1518, B*3801-02, B*3901 , B*3902, B*3903-04, B*4801_02, B*7301, B*2701_〇8 B58: B*1516, B*1517, B*5701, B*5702, B58 B62: B*4601 B52, B*1501 (B62), B*1502 (B75), B*1513 (B77) The population coverage calculations obtained by different HLA supertype combinations are listed in Table IV(g)$. As used herein, "treatment" or "therapeutic" and grammatical terms refer to any improvement in the consequences of any disease, such as prolonged survival, reduced morbidity and/or reduced side effects as a by-product of alternative therapeutic forms; It is easy to understand in the technology that m is not a therapeutic requirement, but it is preferred. The cause (eg, mouse or rat) has a transgenic base = =, ': the transgenic gene is in the pre-natal (eg, embryonic) = or animal ancestor. "Transgenic lean gene" is a DNA within the genome of a cell of a genetic animal J42769.doc -39- 201021828 As used herein, a HLA or cellular immune response "vaccine" is a combination comprising or encoding one or more peptides of the invention. Things. There are various embodiments of such vaccines such as a mixture of one or more individual peptides; one or more peptides of the invention comprising a plurality of epitopes; or a nucleic acid encoding such individual peptides or polymorphisms, eg, A small gene of an epitope peptide. "One or more peptides" may include any integer unit integer of 1 -15 0 or 150 or more, such as at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38 , 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 8〇, 85, 9〇, %, ι〇〇, 1 〇5, 110, 115, 120, 125, 130, 135, 140, 145 or 150 or more than 15 peptides of the invention. The peptide or polypeptide may optionally be modified, such as by lipidation, to add a targeting sequence or other sequence. The HLA-like peptides of the present invention may be mixed or linked with a π-type HLA peptide to help activate cytotoxic lymphocytes and helper T lymphocytes. HLA vaccines may also comprise transiently labeled antigen-presenting cells, such as dendritic cells. The term "variant" refers to a molecule that exhibits variation with the type or normal type, such as in the corresponding position of the specifically described protein (eg, the 58P1D12 protein shown in Figure 1) and the right one, with multiple Proteins of different amino acid residues. The analog is a variant protein. Interview η丄* n 〇 Ά ^ Example. Potentially isomorphic and mono-nucleotide polymorphisms (other examples of SNm variants. The "58P1D12-related protein 皙6" 丄丄 «poor white shell" of the present invention includes the proteins specifically identified herein. And can be used in accordance with the methods described in this article or in the technique 142769.doc 201021828, which is easy to use, to separate/produce and characterize the dual gene variants and conservative substitution variants without improper experimentation. And analogs and homologs. Also included are fusion proteins that combine portions of different 58P1D12 proteins or fragments thereof, and fusion proteins of 58P1D12 proteins and heterologous polypeptides. These 58P1D12 proteins are collectively referred to as 58P1D12-related proteins, proteins of the invention or 58P1D12. The term "58P1D12 related protein" means having 4, 5, 6, 7, 8, 9, 10, 11' 12, 13, 14, 15, 16, 17, 18 ' 19, 20, 21, 22, 23, 24, 25 or more amino acid methic acid polypeptide fragments or 58P1D12 protein sequences; or having at least 3, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100 , 105, 110, 115 , 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 273, 300 or more than 300 amino acids Polypeptide fragment or 58P1D12 protein sequence. II.) 58P1D12 polynucleotide _ an aspect of the invention provides a polynucleus that is identical or complementary to, preferably in isolated form, all or part of the 58P1D12 gene, mRNA and/or coding sequence Glycosylates, including polynucleotides encoding 58P1D12-related proteins and fragments thereof, purine, RNA, DNA/RNA hybrids and related molecules; polynucleotides or oligonucleosides complementary to the 58P1D12 gene or mRNA sequence or a portion thereof An acid; and a polynucleotide or a nucleotide (referred to as "58P1D12 polynucleotide") that hybridizes to the 58P1D12 gene, mRNA, or hybridizes to a polynucleotide encoding 58P1D12. In all cases mentioned in this section, T can also be U in Figure 1. 142769.doc -41· 201021828 Examples of 58P1D12 polynucleotides include: 58P1D12 polynucleus having a sequence as shown in Figure 1 (as shown in Figure 1, wherein T is a nucleotide sequence of 58P1D12 of U) Glycosyl acid; at least 10 such nucleus acids having a polynucleotide of the sequence shown in Figure 1; or at least 1 having a sequence as shown in Figure 1, wherein T is a polynucleotide of U One contiguous nuclear bitter acid. Polynucleotides encoding a relatively long portion of the 58P 1D12 protein are also within the scope of the invention. For example, encoding a 58P1D12 protein or "variant" of about amino acid guanidine (or 2 or 30 or 40, etc.) as shown in Figure 1 or Figure 3 to about amino acid 20 (or 30 or 40 or Polynucleotides of 50, etc. can be produced by a variety of techniques well known in the art. Such polynucleotide fragments may include any portion of the 58P1D12 sequence as shown in Figure 1. ΙΙ·Α·) 5 Use of the 8P1D12 polynucleotide ΙΙ.Α.1 Monitoring genetic abnormalities The polynucleotides of the preceding paragraphs have a variety of different specific uses. The human 58P1D12 gene is located in the chromosomal location set forth in the example entitled "Chromosome localization of 58P1D12". For example, since the 58ρ丨D丨2 gene is localized in this chromosome, a polynucleotide encoding a different region of the 58P1D12 protein is used to discriminate the cytogenetic abnormality at the chromosomal location, such as an abnormality associated with various cancers. . In some genes, multiple chromosomal abnormalities (including rearrangements) have been identified as frequently occurring cytogenetic abnormalities in many different cancers (see, for example, Krajinovic et al., Mutat. Res. 382(3-4): 81-83 (1998) Johansson et al., Blood 86 (1〇). 3905-3914 (1995) and Finger et al., Ρ·Ν. AS 85(23): 9158- 9162 (1988)). Thus, the poly(142769.doc-42-201021828) nucleotide encoding a specific region of the 58P1D12 protein provides a novel tool that can provide greater accuracy than previously possible, which will encode a region of 58P1D12 A genetic abnormality that leads to a malignant phenotype is depicted. In such cases, such polynucleotides meet the requirements of the art for expanding the sensitivity of chromosome screening in order to identify more subtle and unusual chromosomal abnormalities (see, for example, Evans et al., Am. J. Obstet. Gynecol 171(4): 1055-1057 (1994)). Furthermore, since 58P1D12 has been shown to be highly expressed in ovarian cancer and other cancers, the 58P1D12 polynucleotide can be used in a method of assessing the state of the 58p1D12 gene product in normal tissues versus cancerous tissues. Polynucleotides encoding a specific region of the 58P1D12 protein are typically used to assess whether there is a change in a particular region of the 58P1D12 gene, such as a region containing one or more motifs (such as deletions, insertions, point mutations, or changes in antigenic reduction) ). Exemplary assays include RT_PCR assays and single-strand conformation polymorphism (SSCP) analyses (see, eg, Marrogi et al., J. Cutan. Pathol. φ 26 (8): 369-378 (1999)). Polynucleotides encoding specific regions of the protein examine such regions within the protein. II.A.2 antisense embodiment

Other nucleic acid related embodiments specifically contemplated by the invention disclosed herein are genomic DNA, cDNA, ribozymes, and antisense molecules, as well as nucleic acid molecules based on alternative backbones or including alternatives (whether obtained from natural sources or synthesized) And includes molecules capable of inhibiting the expression of RNA or protein of 58P1D12. For example, an antisense molecule can be an RNA or other molecule, including a peptide nucleic acid (PNA) or a non-nucleic acid molecule that specifically binds dna or RNA 142769.doc-43-201021828 in a base-pair dependent manner, such as a thio scale. Sour vinegar derivatives. Those skilled in the art can readily obtain nucleic acid molecules of these classes using the 58P1D12 polynucleotides and polynucleotide sequences disclosed herein. Antisense technology requires the administration of an exogenous oligonucleotide that binds to a target polynucleotide located within the cell. The term "antisense" means that the oligonucleotides are complementary to their intracellular targets (e.g., 58P1D12). See, for example, Jack Cohen, Ol igodeoxy nucleotides, Antisense Inhibitors of Gene

Expression, CRC Press, 1989; and Synthesis 1:1-5 (1988). The 58P1D12 antisense oligonucleotide of the present invention includes a derivative exhibiting enhanced cancer cell growth inhibitory action, such as an S-oligonucleotide (phosphorophosphoric acid derivative or S-oligo, see Jack Cohen, supra). S-oligo (nucleoside thiophosphoric acid vinegar) is an electronic analog of oligonucleotide (〇-〇lig〇) in which a non-bridged oxygen atom of an acid-filled group is replaced by a sulfur atom. The S-〇lig〇 of the present invention can be produced by treating O-oligo with a sulfur transfer reagent 3H-1,2-benzodithiol-3-one-1,1-dioxide. See, for example, Iyer, R. P. et al., J 〇rg Chem. 55: 4693-4698 (1990); and Iyer, R. Ρ et al., j. Am Chem. Soc. 112: 1253-1254 (1 990 ). Other 58P]D12 antisense raised nucleotides of the invention include morpholino antisense sulphuric acid known in the art (see, for example, Partridge et al, 1996, Antisense & Nucleic Acid Drug Development 6: 169- 175). The 58P1D12 antisense oligonucleotide of the present invention may generally be RNA or DNA which is complementary to and stably hybridizes with the 58P1D12 genomic sequence or the first 100 5, codon or end 〇〇 3, codons of the corresponding mRNA. There is no need for absolute complementarity, but South complementarity is better. The use of oligonucleotides complementary to this region allows selective hybridization with 142769.doc -44 - 201021828 58P1D12 mRNA without selective hybridization to mRNAs of other regulatory subunits that define protein kinases. In one embodiment, the 58P1D12 antisense nucleus nucleotide of the present invention is a 15-mer to 〇 mer fragment of the antisense DNA molecule and has a sequence which hybridizes to the 58P1D12 mRNA. The 58P1D12 antisense nucleotide is optionally an oligo-nucleotide complementary to the region of the first 5 , 5, codon or the last 10 3' codons of 58P1D12. Alternatively, the antisense molecule is modified to inhibit 58plD12 expression using a ribozyme, see, for example, L. A. Couture & D. T. Stinchcomb; Trends Genet 12: 510-515 (1996). II.A.3 Primers and Primers Other specific examples of such nucleotides of the invention include primers and primer pairs that permit specific amplification of the polynucleotide of the invention or any particular portion thereof, and the invention A probe that selectively or specifically hybridizes to a nucleic acid molecule or any portion thereof. The probe may be labeled with a detectable label such as a radioisotope, a fluorescent compound, a bioluminescent compound, a chemiluminescent compound, a metal chelating agent or an enzyme. These probes and primers are used to detect the presence or absence of 58p1D12 polynucleic acid in the sample and as a means of detecting cells expressing the 58P1Di2 protein. Examples of such probes include polypeptides comprising all or part of the human 58P1D12 cDNA sequence as shown in Figure 1. Examples of primer pairs capable of specifically amplifying 58 piD12 mRNA are also described in the examples. As will be appreciated by those skilled in the art, a wide variety of different primers and probes can be made based on the sequences provided herein and can be effectively used to amplify and/or detect 58P1D12 mRNA. The 58P1D12 polynucleotide of the present invention can be used for various purposes, including (but not limited to, 142769.doc • 45·201021828) as a probe and primer for amplifying and/or detecting the 58P1D12 gene, mRNA or a fragment thereof; An agent for the diagnosis and/or prognosis of cancer and other cancers; a coding sequence capable of directing the expression of a 58P1D12 polypeptide; a tool for regulating or inhibiting the expression of the 58P1D12 gene and/or translation of a 58P1D12 transcript; and as a therapeutic agent . The invention encompasses the use of any of the probes described herein to identify and isolate a 58P1D12 or 58P1D12-related nucleic acid sequence from a naturally occurring source, such as a human or other mammal, and to use the isolated nucleic acid sequence itself All or most of the sequences seen in the probe. II.A.4 Isolation of Nucleic Acid Molecules Encoding 58P1D12 The 58P1D12 cDNA sequence described herein enables the isolation of other polynucleotides encoding the 58P1D12 gene product and enables the encoding of the 5 8P1D12 gene product homolog or the 58P1D12 gene product. The splicing isoforms, the dual gene variants and the mutant form of the polynucleotide, and the polynucleotide encoding the analog of the 58P1D12 related protein are isolated. A variety of molecular selection methods that can be used to isolate full-length cDNA encoding the 58P1D12 gene are well known (see, for example, Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, New York, 1989; Current Protocols In Molecular Biology. Ausubel et al., Wiley and Sons, 1 995). For example, lambda phage selection methods can be conveniently employed using commercially available colonization systems (e.g., Lambda ZAP Express, Stratagene). A phage-pure line containing the 58P1D12 gene cDNA can be identified by detection with the labeled 58P1D1 2 cDNA or a fragment thereof. For example, in an embodiment, a 58P1D12 cDNA (eg, Figure 1) or a portion thereof can be synthesized and used as a probe to obtain overlapping and full-length cDNA corresponding to the 58p1D12 gene. The 58P1D12 gene itself can be isolated by screening the genomic dnA library, bacterial artificial chromosome library (BAC), yeast artificial chromosome library (YAC) and the like with a 58P1D12 DNA probe or primer. II. A.5 recombinant nucleic acid molecule and Host-Vector System The invention also provides a recombinant DNA or RNA molecule comprising a 58P1D12 polynucleotide, a fragment thereof, an analog or a homologue, including but not limited to, a bacterium, a plastid, a phage ( Phagemid), viscous, YAC, BAC, and a variety of viral and non-viral vectors well known in the art, and cells transformed or transfected with such recombinant DNA or RNA molecules. Methods for producing such molecules are well known (see For example, Sambrook et al., 1989, supra. The invention further provides a host-vector system comprising a 58P1D12 polynucleotide, fragment, analog or homolog thereof in a suitable prokaryotic or eukaryotic host cell. Recombinant DNA molecules. Examples of suitable eukaryotic host cells include yeast cells, plant cells or animal cells, such as mammalian cells or insect cells (e.g., cells that can be infected with baculovirus, such as Sf9 or HighFive cells). Examples of mammalian cells include various ovarian cancer cell lines (such as OVCAR-5 and CaOV-3, other transfectable or transducible ovarian cancer cell lines), primary cells (PrEC), and various mammals commonly used to express recombinant proteins. Cells (eg, COS, CHO, 293, 293T cells). More specifically, a polynucleotide comprising a coding sequence for 58P1D12 or a fragment, analog or homolog thereof can be used in order to use the commonly used and widely used in the art. .doc • 47· 201021828 A number of host-vector systems are known to produce 58P1D12 proteins or fragments thereof. A variety of host-vector systems suitable for the expression of 58P1D12 proteins or fragments thereof can be obtained, see, for example, Sambrook et al., 1 989, supra; Current Protocols in Molecular Biology, 1995, supra. Preferred carriers for mammalian performance include (but are not limited to pcDNA 3.1 myc-His-tag (Invitrogen) and the retroviral vector pSR tkneo (Muller et al, 1991, MCB 1 1:1785). Using these expression vectors, 58ρ1〇ΐ2 can be used in several ovarian cancers and non- Ovarian cancer cell lines (including, for example, 293, 293T, rat-1, NIH 3T3, and TsuPrl) are expressed. The host-vector system of the invention can be used to prepare 58P1D12 protein or a fragment thereof. These host-vector systems can be used to study the functional properties of 58P1D12 and 58P1D12 mutants or analogs. Recombinant human 58P1D1 2 protein or an analog or homolog or fragment thereof can be prepared by mammalian cells transfected with a construct encoding a 58P1D12-related nucleotide. For example, '293T cells can be transfected with a plastid encoding the 5 8P1D12 or a fragment, analog or homolog thereof, and the 58P1D12-related protein is expressed in 293T cells, and affinity is used using standard purification methods (eg, using an anti-58P1D12 antibody). Purification) Recombinant 58p1D12 protein was isolated. In another embodiment, the 58P1D12 coding sequence is sub-selected into the retroviral vector pSRcxMSVtkneo and used to infect a variety of mammalian cell lines, such as NIH 3T3, TsuPrl, 293, and rat-Ι, to establish a 58P1D12 expression cell line. A variety of other performance systems well known in the art can also be used. The expression construct of the leader peptide 142769.doc -48-201021828 can be used in the same frame as the 58P1D12 coding sequence for use in the gentleman tongue for the secretory form of the 58P1D12 protein. As described herein, genetic code redundancy allows the 58 pim2 gene sequence to mutate. In particular, it is known in the art that a particular host species typically has a particular codon preference, and thus the disclosed sequences can be adapted to the desired host. For example, a rare codon of a preferred analog codon sequence (i.e., a codon having a frequency of use of less than about 2% in a known sequence of a desired host) is typically replaced with a higher frequency codon. For example, a particular type of codon preference can be calculated by using a codon usage table available on the Internet (such as URL dna affrc g〇 jp/~ nakamura/c〇d〇n.html). Other sequence modifications are known to enhance the performance of proteins in cellular hosts. Such sequence modifications include the elimination of a sequence encoding a false polyadenylation signal, an exon/intron splice site signal, a transposon-like repeat, and/or other such well-characterized sequences that are detrimental to gene expression. . The _GC content of the sequence is adjusted to the average degree of a given cellular host as calculated by reference to known genes expressed in the host cell. If feasible, the sequence is modified to avoid the expected secondary mRNA hairpin structure. Other useful modifications include adding a translation start consensus sequence at the beginning of an open reading frame, such as Kozak.

Mol. Cell Biol., 9: 5073-5080 (1989). Those skilled in the art should be aware that the general rule that eukaryotic ribosomes initiate translation at only the proximal AUG codon is only abolished under rare conditions (see, for example, Kozak PNAS 92(7): 2662-2666, (1995). And Kozak NAR 15(20): 8125-8148 (1987)) 〇 142769.doc -49- 201021828 III.) 58P1D12 Related Proteins Another aspect of the invention provides a 58P1D12 related protein. A specific embodiment of the 58P1D12 protein comprises a polypeptide having all or part of the human 58P1D12 amino acid sequence as shown in Figure 1 (preferably Figure 1A). Alternatively, an embodiment of the 58P1D12 protein comprises a variant, homolog or analog polypeptide having a variation in the 58卩1〇12 amino acid sequence shown in Figure 1. Examples of the 5P1D12 polypeptide include: a 58P1D12 polypeptide having the sequence shown in Figure 1; a peptide encoded by a polynucleotide sequence of 58P1D12 in which τ is u as shown in Figure 1; At least 10 b-specific nucleotides of the polypeptide of the sequence shown; or at least one of the contiguous peptides encoded by a polynucleotide having a sequence of T as shown in Figure 1 . Amino acid abbreviations are provided in Table II. Conservative amino acid substitutions can often be made in proteins without altering the conformation or function of the protein. The protein of the present invention may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 conservative substitutions. Embodiments of the invention disclosed herein include various variants or analogs of the 58P1D12 protein accepted in the art, such as polypeptides having amino acid hydrazine insertions, deletions, and substitutions. The 58P1D12 variant can be prepared using methods known in the art, such as site-directed mutagenesis, alanine scanning, and PCR mutation induction. Site-directed mutagenesis can be performed on selected DNA (Carter et al. 'Nucl. Acids Res., 13:433 1 (1986); Zoller et al., Nucl. Acids Res, 10:6487 (1987)), cassette Mutation induction (Wells et al, Gene, 34: 315 (1985)), restriction-selective mutation induction (Wells Specialist' Philos. Trans. r. s〇c. London SerA, 317:415 142769.doc •50· 201021828 ( 1986)) or other known techniques 'to prepare a 58 piDi2 variant dNA. One or more amino acids can also be identified along a contiguous sequence involving a particular biological activity, such as a protein-protein interaction, using a scanning amino acid assay. Preferably, the scanning amino acid is a relatively small neutral amino acid. Such amino acids include alanine, glycine, serine and cysteine. Alanine is generally the preferred scanning amino acid in this group because it eliminates the side chain beyond the beta carbon and is unlikely to alter the backbone configuration of the variant. Alanine is generally preferred because it is the most common amino acid. Furthermore, it is often present in the buried and exposed locations (Creighton, The Proteins, (w.h. Freeman & Co., N.Y.); Chothia, J. Mol. Bi〇l., i5〇: i (1976)). If the alanine substitution does not produce an appropriate amount of variant, an isosteric amino acid can be used. As defined herein, a distinguishing characteristic of a 58P1D12 variant, analog or homolog is that it has at least one epitope that is "cross-reactive" with the 58P1D12 protein having the amino acid sequence of the map. As used in this sentence, "cross-reactivity" means that the antibody or T cell that specifically binds to the 58piD12 variant also specifically binds to the 58P1D12 protein having the amino acid sequence set forth in Figure 1. When the polypeptide no longer contains any epitope that can be recognized by an antibody or tau cell that specifically binds to the initial 58P1D12 protein, the polypeptide is not a variant of the protein shown in Figure 1. Those skilled in the art will appreciate that antibodies that recognize proteins that bind to different size epitopes and a group of about four or five contiguous or non-contiguous amino acids are considered typical of the amino acid in the minimal epitope. number. See for example 1^ to 1 etc.

Human ' J. Immunol 2000 165 (12): 6949-6955; Hebbes et al, Mol Immunol (1989) 26(9): 865-73; Schwartz et al, J 142769.doc -51- 201021828

Immunol (1985) 135(4): 2598-608 ° Other 58P1D12-related protein variants share 70%, 75% '80%, 85%, 90%, 95% of the amino acid sequence of Figure 1 or its fragment Or 95% or more similarity. Another particular class of 12 protein variants or analogs comprises one or more of the 58P1D12 biological motifs described herein or currently known in the art. Thus, the invention encompasses analogs of a 58p iD12 fragment (nucleic acid or amino acid) having altered functional (e.g., immunogenic) properties relative to the starting fragment. It will be appreciated that a motif that is or will be part of this technology is intended for use in the nucleic acid or amino acid sequence of Figure 1. As described herein, embodiments of the invention include polypeptides comprising an intact amino acid sequence that is less than the 58P1D12 protein shown in Figure 1. By way of example, a representative embodiment of the invention comprises any of four, five, six, seven, eight, nine, one, eleven, eleven, twelve having the 58P1D12 protein shown in FIG. , 13, 14, 15 or more peptides/proteins adjacent to the amino acid. The 58P1D12 related protein can be produced using standard peptide synthesis techniques well known in the art or using chemical cleavage methods. Alternatively, a recombinant method can be used to generate a nucleic acid molecule encoding a 5 8P1D12 related protein. In one embodiment the 'nucleic acid molecule provides a method of producing a defined fragment of a 58P1D12 protein (or variant, homolog or analog thereof). III.A.) Proteins with Primers Examples Other illustrative embodiments of the invention disclosed herein include 58P1D12 polypeptides. The 58P1D12 polypeptides comprise one or more of the 58P1D12 polypeptide sequences as listed in Figure 1. Amino acid residues in the biological motifs within 142769.doc -52- 201021828. A variety of motifs are known in the art' and the presence of the temple base in the protein can be assessed by a number of publicly available internet sites such as BIMAS. A motif having a subsequence of all 58P1D12 variant proteins has been previously disclosed. Based on pfam search (see URL address pfam.wustl.edu/), table iv(h) lists several frequently occurring primitives. The rows in Table IV(h) are listed: (1) abbreviations for the base name; (2) the consistency percentage found between different members of the primitive family; (3) the name or description of the primitive; 4) The most common function; if the primitive is related to the location, it includes location information. Since the above 58P1D12 motif is observed to be associated with growth dysregulation and because 58P1D12 is overexpressed in certain cancers (see, eg, Table j), polypeptides containing one or more of the above 58P1D12 motifs can be used to elucidate specific features of malignant phenotypes. . For example, tic-kinase kinase c, c AMP and camp-dependent protein kinases and protein kinase c are enzymes known to be associated with the appearance of g of the malignant phenotype (see, eg, Chen et al, Lab Invest., 78(2): 165-174 (1998), Gaiddon et al, Endocrinology 136(10): 4331-4338 (1995), Hall et al, Nucleic Acids Research 24(6): 1119-1126 (1996); Peterziel et al, Oncogene 18 ( 46): 6322-6329 (1999); and O'Brian, Oncol. Rep. 5(2): 305-309 (1998)). Furthermore, both glycosylation and myristylation are protein modifications that are also associated with cancer and cancer development (see, for example, Dennis et al., Biochem

Biophys. Acta 1473(1): 21-34 (1999); Raju et al., Exp. Cell Res. 235(1): 145-154 (1997)). Amidoxime is another protein modification that is also associated with cancer and cancer 142769.doc • 53- 201021828 (see, eg, Treston et al., Natl. Cancer Inst. Monogr. (13): 169-175 (1992)) . In another embodiment, the protein of the invention comprises one or more immunoreactive epitopes (such as the peptides disclosed above) identified according to methods accepted in the art. A specific algorithm can be used to determine the CTL epitope to identify a specific HLA allele within the 58P1D12 protein (eg, Table IV; EpimatrixTM & EpimerTM, Brown University, URL brown.edu/Research/TB-HIV_Lab/epimatrix/epimatrix.html; And BIMAS, URL bimas. dcrt.nih.gov/.) The best combination of peptides. In addition, methods for identifying peptides having sufficient binding affinity for HLA molecules and associated with immunogenic epitopes are well known in the art and can be performed without undue experimentation. Furthermore, methods for identifying peptides that are immunogenic epitopes are well known in the art and can be performed in vitro or in vivo without undue experimentation. The principle of forming analogs of such epitopes to modulate immunogenicity is also known in the art. For example, one principle may begin with an epitope having a CTL or HTL motif (see, for example, class I HLA and class II HLA motifs/supercells of Table IV). An epitope is modeled by substituting an amino acid at a particular position and replacing it with another amino acid designated for that position. For example, based on the residues defined in Table IV, the deleterious residue can be substituted with any other advantageous residue, such as a preferred residue; the poor residue can be substituted with a preferred residue; or the original Preferred residues present are substituted with another preferred residue. Substitutions can occur at a single anchor or other position in the peptide; see, for example, Table IV. 142769.doc -54· 201021828 Many references reflect techniques related to the identification and production of antigenic determinants and their analogues in proteins. See, for example, Chesnut et al., WO 97/33602; Sette, Immunogenetics 1999 50(3-4): 201-212; Sette et al, J. Immunol. 2001 166(2): 1389-1397;

Sidney et al, Hum. Immunol. 1997 58(1): 12-20; Kondo et al, Immunogenetics 1997 45(4): 249-258; Sidney et al, J. Immunol. 1996 157(8)·· 3480- 90; and Falk et al, Nature 351: 290-6 (1991); Hunt et al, Science 255: 1261-3 (1992); • Parker et al, J. Immunol. 149:3580-7 (1992); Parker Et al., J. Immunol. 152: 163-75 (1994)); Kast et al., 1994 152(8): 3904-12; Borras-Cuesta et al., Hum. Immunol. 2000 61(3): 266-278 Alexander et al, J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al, PMID: 7895164, UI: 95202582; O'Sullivan et al, J. Immunol. 1991 147 (8 ): 2663-2669 ; Alexander et al., Immunity 1994

1(9): 751-761; and Alexander et al., Immunol. Res. 1998 18(2): 79-92. i Related embodiments of the invention include different primitives listed in Tables IV(a), IV(b), IV(c), IV(d), and IV(h) and/or one or more prior A polymorphism of a combination of a predicted CTL epitope and/or one or more T cell binding motifs known in the art is disclosed. The preferred embodiment does not contain insertions, deletions or substitutions within the polymorphic motif or within the interventional sequence. Furthermore, embodiments comprising a plurality of N-terminal and/or C-terminal amino acid residues on either side of the motifs may be desirable (e.g., including a larger portion of the polypeptide structure in which the motif is located 142769.doc) -55- 201021828 points). The number of N-terminal and/or C-terminal amino acid residues located on either side of the motif is typically from about 1 to about 1 amino acid residues, preferably from 5 to about 5 amino acid residues. Between the bases. The 58P1D12 related protein can be present in a variety of forms, preferably in isolated form. The purified 58P1D12 protein molecule is substantially free of other proteins or molecules that interfere with the binding of 58P1D12 to antibodies, tau cells or other ligands. The nature and extent of the separation and purification will depend on the intended use. Examples of 5 8卩1012 related proteins include purified 58?1〇12 related proteins and functional, soluble 58P1D12 related proteins. In one embodiment, the functional, soluble 58P1D12 protein or fragment thereof retains the ability to bind to an antibody, T cell or other ligand. The invention also provides a 58P1D12 protein comprising a biologically active fragment of the 58P1D12 amino acid sequence as shown in Figure 1. Such proteins exhibit the properties of the starting 58P1D12 protein 'such as the ability to elicit an antibody that specifically binds to an epitope associated with the initiation of the 58P1D12 protein; the ability to bind to such antibodies; the ability to induce HTL or CTL activation; and / Or the ability to specifically bind to the HTL of the starting protein or the ability to recognize. 3 The 5 8 P1D12-related polymorphism with a particularly desirable structure can be predicted using a variety of analytical techniques well known in the art and/or Identification, such analysis techniques include, for example, Chou-Fasman, Garnier-Robson 'Kyte-

Doohttle, Eisenberg, Karplus-Schultz or Jameson-Wolf analytical methods or methods based on immunogenicity. Fragments containing such structures are particularly useful for generating subunit specific anti-58P1Di2 antibodies or T cells or for identifying cytokines that bind to 5 8P1D12. For example, the method of H〇pp, 142769.doc -56-201021828 Τ·Ρ· and Woods, KR·, 1981, Proc. Natl. Acad. Sci. USA 78: 3 824-3 828 can be used to produce hydrophilicity. Curve and identify immunogenic peptide fragments. The hydrophobicity profile can be generated and the immunogenic peptide fragments can be identified using the method of Kyte, J. and Doolittle, R. F., 1982, J. Mol. Biol. 15 7: 105-132. The percent residue (%) curve can be generated and the immunogenic peptide fragments can be identified using the method of Janin J., 1979, Nature 277: 491-492. The average flexibility curve can be generated and the immunogenic peptide fragments can be identified using the method of Bhaskaran R., Ponnuswamy P.K., 1988, Int. J. Pept. Protein Res. 32:242-255. The β-turn curve can be generated and the immunogenic peptide fragment can be identified using the method of Deleage, G., Roux B., 1987, Protein Engineering 1: 289-294. A specific algorithm can be used to determine the CTL epitope to identify peptides within the 58P1D12 protein that are optimally associated with a particular HLA allele, such as BIMAS and S YFPEITHI. To illustrate this, peptide epitopes in 58P1D12 presented in the context of human class I MHC molecules (e.g., HLA-A1, Α2, A3, All, Α24, Β7, and B35) are predicted. For the specific reference, the complete amino acid sequence of the relevant part of the 58P1D12 protein and other variants (ie, for the class I HLA prediction, there are 9 flanking residues on either side of the point mutation or exon junction) And for class II HLA predictions, there are 14 flanking residues on either side of the point mutation or exon junction of the variant.) HLA peptide primitive search algorithm as seen in the section on input bioinformatics and molecular analysis. in. The HLA peptide primitive search algorithm was developed by Dr. Ken Parker based on the binding of specific peptide sequences to the grooves of class I HLA molecules (especially HLA-A2) 142769.doc -57- 201021828 (see for example Falk et al., Nature 351: 290-6 (1991); Hunt et al, Science 255: 1261-3 (1992); Parker et al, J. Immunol. 149: 3580-7 (1992); Parker et al, J. Immunol. 163-75 (1994)). This algorithm allows for the localization and categorization of 8-mer, 9-mer and 10-mer peptides in intact protein sequences that are predicted to bind HLA-A2 and various other class I HLA molecules. A variety of class I HLA binding peptides are 8-mer, 9-mer, 1 Ο-mer or 11-mer. For example, for class I HLA-A2, the epitope preferably contains leucine (L) or guanidine thiocyanate (M) at position 2 and valerine (V) or leucine at the C-terminus ( L) (see for example Parker et al, J. Immunol. 149: 3580-7 (1992)). The selection of 58P1D12 predicted binding peptides has been demonstrated. At 37 ° C, at pH 6.5, the binding score corresponds to an estimate of the dissociation half-life of the peptide-containing complex. It is predicted that the peptide with the highest binding score binds most closely to the class I HL A on the cell surface for the longest time and thus represents the optimal immunogenic target for T cell recognition. The actual binding of the peptide to the HLA allele can be assessed by stabilizing the performance of HL A on the antigen-defective cell line T2 (see, for example, Xue et al.

* G

Prostate 30: 73-8 (1997) and Peshwa et al., Prostate 36: 129-38 (1998)). The immunogenicity of a specific peptide can be assessed in vitro by stimulating CD8+ cytotoxic T lymphocytes (CTL) in the presence of antigen presenting cells such as dendritic cells. It is understood that Class I or Class II HLA motifs (such as those listed in Table IV) are predicted by BIMAS sites, EpimerTM & EpimatrixTM sites or by the technology available or to be part of this technology. Each of the epitopes specified is intended to be "applied" to the 58P1D12 protein of the present invention. As used hereinafter, "application" means that the 58P1D12 protein can be evaluated by a method known to those skilled in the art, visually or by a computer-based pattern finding method. 58P1D12 protein having a class I HLA motif and having each subsequence of 8, 9, 10 or 11 amino acid residues or a subsequence having a class II HLA motif and having 9 or more amino acid residues In the examples described in the following examples, 58P1D12 can be conveniently placed on a commercially available expression vector (such as having a C-terminal 6XHis and MYC marker). The CMV-driven expression vector (pCDNA3.1/mycHIS, Invitrogen, or Tag5, GenHunter Corporation, Nashville TN) encoding 58P1D12 was expressed in transfected cells (such as 293T cells). The Tag5 vector provides an IgGK secretion signal, and the use of the igGK secretion signal can contribute to the production of the secreted 58P1D12 protein in the transfected cells. For example, the HIS marker 58P1D12 secreted in the culture medium can be purified using a standard column using a standard technique. ❹ III.C.) Modification of 58P1D12 Related Proteins Modifications of 58P1D12 related proteins, such as covalent modifications, are included within the scope of the present invention. One type of covalent modification involves reacting a target amino acid residue of a 58P1D12 polypeptide with an organic derivatizing agent capable of reacting with a selected side chain or an N-terminal residue or a c-terminal residue of the 58P1D12 protein. Another type of covalent modification of the 58P1D12 polypeptide encompassed within the scope of the invention comprises altering the native glycosylation pattern of the protein of the invention. Another type of covalent modification of 58P1D12 is disclosed in U.S. Patent Nos. 4,640,835, 142, 769, doc-59-2010, 218, 218, 4, 496, 689, 4, 301, 144, 4, 670, 417, 4, 791, 192, or 4, 179, 337. This allows the 58?1?12 polypeptide to be linked to one of a variety of non-proteinaceous polymers such as polyethylene glycol (pEG), polypropylene glycol or polyalkylene oxide. The 58P1D12 associated protein of the invention may also be modified to form a fungal molecule comprising 58P1D12 fused to another heterologous polypeptide or amino acid sequence. The silvery molecule can be synthesized chemically or recombinantly. A chimeric molecule can have a protein of the invention fused to another tumor associated antigen or fragment thereof. Alternatively, the protein of the invention may comprise a fusion of a fragment of the 58P1D12 sequence (amino acid or nucleic acid) such that the molecule formed is not directly related to the amino acid or nucleic acid sequence shown in Figure 1 over its entire length. source. The chimeric molecule may comprise a plurality of identical subsequences of 58P1D12. The chimeric molecule may comprise a fusion of a 58P1D12 associated protein with a polyhistidine epitope tag (which provides an immobilized nickel selective binding epitope), a cytokine or a growth factor. The epitope tag is typically located at the amino terminus or slow terminus of the 58 piD12 protein. In an alternate embodiment, the agonist molecule may comprise a fusion of a 58P1D12 associated protein with a particular region of an immunoglobulin or immunoglobulin. For the bivalent form of the chimeric molecule (also known as "immunoadhesin"), this fusion can be made with the fc region of the IgG molecule. Preferably, the Ig fusion comprises replacing at least one variable region within the Ig molecule with a soluble (transmembrane domain deletion or inactivation) form of the 58P1D12 polypeptide. In a preferred embodiment, the immunoglobulin fusion comprises igG] [hinge region of the molecule, cH2 region and CH3 region] or the hinge region, CH1 region, CH2 region and CH3 region of the IgGI molecule. For the preparation of immunoglobulin fusions, see, e.g., U.S. Patent No. 5,428,130, issued June 27, 1972, to s. III.D.) Use of 58P1D12 Related Proteins The proteins of the present invention have a variety of different specific uses. Since 58 piD12 is highly expressed in ovarian cancer and other cancers, the 58P1D12-related protein can be used in a method for assessing the state of the 58P1D12 gene product in normal tissues versus cancerous tissues, thereby indicating a malignant phenotype. Polypeptides from specific regions of the 58P1D12 protein are typically used to assess the presence or absence of perturbations (such as deletions, insertions, point mutations, etc.) in those regions, such as regions containing one or more motifs. To assess the characteristics of this region in normal tissue versus cancerous tissue or to induce an immune response against an epitope, exemplary assays use antibodies or tau cells from the 58P1D12-related protein, which contain one or more inclusions. An amino acid residue of a biological motif within the 58 piD12 polypeptide sequence. Alternatively, the 58P1D12-related protein screen containing the amino acid residue of one or more biological motifs in the 58P1D12 protein is screened for a factor 0 58P1D12 protein fragment/subsequence that interacts with the region of 58P1D12 and is particularly useful for generating and characterizing structures. Domain-specific antibodies (eg, antibodies that recognize the extracellular or intracellular epitope of the 58P 1D12 protein) are useful for identifying agents or cytokines that bind to 58P1D12 or a particular domain thereof and are suitable for use in a variety of therapeutic and diagnostic settings, These include, but are not limited to, diagnostic assays, cancer vaccines, and methods of preparing such vaccines. A protein encoded by the 58P1D12 gene or an analog, homolog or fragment thereof has a variety of uses including, but not limited to, production of antibodies and ligands for identifying binding to the 142769.doc • 61 · 201021828 5 8P1D12 gene product and others In the method of the agent and the cellular component. Antibodies raised against the 58P1D12 protein or fragment thereof are useful in diagnostic and prognostic assays; in imaging methods controlled by human cancers characterized by 58P1D12 protein expression, such as those listed in Table I(R). Such antibodies can be expressed in an intracellular manner and can be used in methods for treating patients suffering from such cancers. The 58P1D12 related nucleic acid or protein is also used to generate HTL or CTL responses. A variety of immunological assays for the detection of 58P1D12 proteins can be used, including but not limited to various types of radioimmunoassay, enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunofluorescence assay (ELIFA), immunocytochemistry Methods and similar methods: The antibody can be labeled and used as an immunological imaging reagent capable of detecting 58P1D12 expressing cells (e.g., in a radioactive scintigraphy imaging method). As further illustrated herein, the 58P1D12 protein is also particularly useful for the production of cancer seedlings. IV.) 58P1D12 Antibody Another aspect of the invention provides an antibody that binds to a 5 8 P1D12 associated protein. Preferably, the antibody specifically binds to a 58P1D12-related protein and does not bind (or weakly bind) under physiological conditions to a protein or protein that is not a 58P1D12-related protein. Examples of physiological conditions include: 丨 disc salt buffered saline; 2) Tris buffered saline containing 25 mM Tris and 150 mM NaCl; or physiological saline (0.9% NaCl); 4) animal serum, Such as human serum; or 5) any combination of 1) to 4); such reactions are preferably carried out at pH 75 or in the range of pH 7.0 to 8.0 or in the range of pH 6.5 to 8.5. Furthermore, such reactions At 4 ° ° C to 37. (: Between the temperatures of 142769.doc-62.201021828, an antibody that binds to 58P1D12 can bind to a 58P1D12-related protein, such as a homolog or analog thereof. In one embodiment, the invention comprises: 1] comprises a light chain variable region sequence as shown in SEQ ID NO: 18, 21 to 1; 33 or as shown in SEQ. ID NO: 19, 21 to 134, and SEQ. ID NO: An antibody or fragment of the heavy chain variable region sequence shown in the 20th to 146th of the invention; [2] the antibody or fragment of [1], wherein the antibody specifically binds to the 58P1D12 protein (Fig. 1); The antibody or fragment of [1], wherein the antibody inhibits tumor cell migration and invasion; [4] the antibody or fragment of [4], wherein the antibody comprises 21 to 239 of SEQ. ID NO: 18. a light chain sequence as shown in SEQ ID NO: 19, and a heavy chain sequence comprising the sequence shown in SEQ ID NO: 17 from 20 to 203; encoding [ a light chain or heavy chain polynucleotide of an antibody of 丨^ to ^]; [6] a vector comprising the polynucleotide of [5]; [[7] a cell transfected with the vector of [6]; 8] [7] the cell in which the cell Transfection of a polynucleotide comprising a light chain encoding the antibody of [1] to [4] and a polynucleotide encoding a heavy chain of [丨] to the anti-φ body, or comprising the coding [1] to [4] a vector of a polynucleotide of a light chain of an antibody and a vector comprising a polynucleotide encoding a heavy chain of an antibody of [丨] to [4]; [9] a preparation comprising SEQ. ID The light chain variable region sequences of 21st to 133th of NO: 18 or 21st to 134th of SEQ. ID NO: 19 and 20th to 146th of SEq·ID no: 17 Method of antibody or fragment of a heavy chain variable region sequence 'This method comprises: i) cultivating the cells of [7] under conditions which promote expression of the antibody or fragment, and U) isolating antibodies or fragments from such cells , the method of preparing the antibody or fragment; [1] [9], wherein the antibody comprises, for example, 142769.doc-63·201021828 SEQ ID NO: 18, 21 to 239 or SEQ. ID NO: 19 The light chain sequence shown in the 21st to 240th and the heavy chain sequence comprising the sequence shown as 20th to 20th in SEQ. ID NO: 17. The 58P1D12 antibody of the present invention is particularly useful in the diagnosis and development of cancer (see, e.g., for example) diagnostic and prognostic methods. Similarly, such antibodies are useful in the treatment, diagnosis, and/or prognosis of ovarian cancer and other cancers because 5 8P1D12 is also manifested or overexpressed in such other cancers. Further 'intracellularly expressed antibodies (e. g., single chain antibodies) are therapeutically useful for treating cancers that are indicative of 58P1D12, such as advanced or metastatic ovarian cancer or other advanced or metastatic cancer. The invention also provides various immunoassays suitable for detecting and quantifying 58P1D12 and mutant 5 8P1D12 related proteins. Such assays may optionally include one or more 58P1D12 antibodies that recognize and bind to the 58P1Di2-related protein. Such assays can be performed in a variety of immunoassay formats well known in the art including, but not limited to, various types of radioimmunoassays, enzyme-linked immunosorbent assays (EUSA), enzyme-linked immunofluorescence assays (ELIFA), and the like. Verification.免疫 The immunological non-antibody assay of the invention also comprises a tau cell immunogenicity assay (inhibitory or irritating) and a major histocompatibility complex (MHC) binding assay. In addition, the present invention also provides immunoimaging methods capable of detecting ovarian cancer and other cancers exhibiting 581 > 11) 12, including, but not limited to, radioactive scintigraphy imaging methods using labeled 58P1D12 antibodies. These tests are available on the spot for the detection of 58P1D12 cancers (such as Insect Cancer), and the 142769.doc -64 - 201021828 and prognosis. The 58卩1012 antibody can also be used in methods for purifying 58?1012 related proteins and isolating 5<8>P1D12 homologs and related molecules. For example, a method of purifying a 58P1D12-related protein comprises: cultivating a 58P1D12 antibody that has been conjugated to a solid substrate with a lysate or other solution containing a 58P1D12-related protein, while allowing the 58P1D12 antibody to bind to the 58P1D12-related protein; A solid matrix to eliminate impurities; and a 58P1D12 related protein is detached from the coupled antibody. Other uses of the 58P1D12 ® antibodies of the invention include the production of anti-genotype antibodies that mimic the 58P1D12 protein. A variety of methods for preparing antibodies are well known in the art. For example, antibodies can be prepared by immunizing a suitable mammalian host using 58P1D12-related proteins, peptides or fragments in isolated or immunoconjugated form (Antibodies: A Laboratory Manual, CSH Press, Harlow and Lane ed. (1988), Harlow , Antibodies, Cold Spring Harbor © Press, NY (1989)). In addition, a 58P1D12 fusion protein such as the 58P1D12 GST fusion protein can also be used. In a specific embodiment, a GST fusion protein comprising all or a majority of the amino acid sequence of Figure 1 is prepared as an immunogen for the production of a suitable antibody. In another embodiment, a 58P1D12 related protein is synthesized and used as an immunogen. In addition, the naked DNA immunization technique known in the art (with or without the use of purified 58P1D12-related proteins or 58P1D12 cells) produces an immune response against the encoded immunogen (for review, see Donnelly et al.) '1997, Ann. Rev. Immunol. 15: 617-648). 142769.doc -65- 201021828 The amino acid sequence of the 58P1D12 protein as shown in Figure 1 can be analyzed to select a specific region of the 58P1D12 protein to produce antibodies. For example, the hydrophobicity and hydrophilicity analysis of the 58P1D12 amino acid sequence was used to identify the hydrophilic regions in the 58P1D12 structure. The region of the 58P1D12 protein displaying the immunogenic structure, as well as other regions and domains, can be readily identified using a variety of other methods known in the art, such as (:}1011-

Fasman, Garnier-Robson, Kyte-Doolittle, Eisenberg, 1^印11^-8〇1111112 or 11^8〇11-\¥〇1 £ analysis. The hydrophilicity curve can be produced using the method of 11 〇?卩, 1\?. and Woods, K.R., 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. The hydrophobicity curve can be produced using the method of Kyte, J. and Doolittle, R. F., 1982, J. Mol. Biol. 157: 105-132. The percent residue (%) curve can be generated using the method of Janin J., 1979, Nature 277:491-492. The average pullability curve can be generated using the method of Bhaskaran R., Ponnuswamy P.K., 1988, Int. J. Pept. Protein Res. 32:242-255. The beta angle curve can be generated using the method of Deleage, G., Roux B., 1987' Protein Engineering 1: 289-294. Therefore, each of the regions identified by any of the programs or methods is within the scope of the present invention. A preferred method of producing the 58P1D12 antibody is further illustrated by the examples provided herein. Methods of preparing proteins or polypeptides for use as immunogens are well known in the art. Methods of preparing immunogenic conjugates of proteins with vectors such as B S A, KLH or other carrier proteins are also well known in the art. In some cases, direct bonding is used, which uses, for example, a carbodiimide reagent; in other cases, linking reagents (such as those supplied by Pierce Chemical Co., Rockford, IL, 142769.doc-66-201021828) are effective. . As is understood in the art, administration of the 58P1D12 immunogen is typically performed by injection over a suitable period of time and using an appropriate adjuvant. Antibody titers can be measured during the immune time course to determine if sufficient antibodies are formed. The 58P1D12 monoclonal antibody can be prepared by a variety of means well known in the art. For example, it is known to prepare an immortalized cell line secreting a desired monoclonal antibody using a standard fusion tumor technique of Kohler and Milstein or a modification technique for immortalizing an antibody to produce B cells. Screening by immunoassay _ is an immortalized cell line of antibodies required for secretion, wherein the antigen is a 5 8P1D12-related protein. When a suitable immortalized cell culture is identified, the cells can be propagated and antibodies produced from the in vitro culture or ascites. The antibodies or fragments of the invention may also be prepared by recombinant means. The region that specifically binds to the desired region of the 5 8P1D12 protein can also be prepared in the context of a multi-species antibody or complementarity determining region (CDR)-implanted antibody. Humanized or human 58P1D12 antibodies can also be prepared and are preferred for use in therapeutic settings. Methods for humanizing murine antibodies and other non-human antibodies by substituting one or more non-human antibody CDRs into corresponding human antibody sequences are well known (see, for example, Jones et al., 1986, Nature 321: 522-525). Riechmann et al., 1988, Nature 332: 323-327; Verhoeyen et al., 1988, Science 239: 1534-1536). See also Carter et al, 1993, Proc. Natl. Acad. Sci. USA 89: 4285 and Sims et al, 1993, J. Immunol. 151: 2296. Methods for preparing fully human monoclonal antibodies include phage display and transgenic methods (for review, see Vaughan et al., 1998, Nature 142769.doc -67-201021828).

Biotechnology 16: 535-539). Using the large-scale human Ig gene combinatorial library (also known as phage display) to generate fully human 58P1D12 monoclonal antibodies (Griffiths and Hoogenboom, Building an vz7ro immune system: human antibodies from phage display libraries ° in Protein Engineering of Antibody Molecules for Prophylactic and Therapeutic Applications in Man, Clark, M. (ed.), Nottingham Academic, pp. 45-64 (1993); Burton and Barbas, Human Antibodies from combinatorial libraries (ibid., pp. 65-82). Fully human 5 8P1D12 monoclonal antibodies can also be prepared using a transgenic mouse engineered to contain the human immunoglobulin locus, as described in the following documents: PCT Patent Application WO 98/24893, Kucherlapati and Jakobovits, et al. Person, published on December 3, 1997 (see also Jakobovits, 1998, Exp. Opin·Invest. Drugs 7(4): 607-614; US Patent 6,162,963, issued December 19, 2000; November 12, 2000 6,150,584 issued; and 6,114,598 issued on September 5, 2000). This method can avoid phage display technology

A requires in vitro manipulation and can efficiently produce high affinity reliability human W antibodies. The reactivity of the 58P1D12 antibody with the 5 8P1D12-related protein may be established by using a 58P1D12-related protein, a 58P1D12-expressing cell or an extract thereof by various well-known methods including Western blotting, immunoprecipitation, ELISA, and FACS analysis. . The 58P1D12 antibody or fragment thereof can be labeled with a detectable label or ligated to a second molecule. Suitable detectable markers include, but are not limited to, radioisotopes, fluorescent compounds, biological hair 142769.doc -68- 201021828 photocompounds, chemiluminescent compounds, metal chelators or enzymes. Furthermore, bispecific antibodies which are specific for two or more 58P1D12 epitopes can be produced using methods generally known in the art. Homodimeric antibodies can also be produced by cross-linking techniques known in the art (e.g., Wolff et al, Cancer Res. 53: 2560-2565). In one embodiment, the invention provides a monoclonal antibody produced by a fusion tumor that has been labeled Ha8-4c4.1, which was delivered to the United States on August 5, 2008 (via Federal Express) The American Type Culture Collection (ATCC) (mail 1549, Manassas, VA 20108) and the registration number PTA-9404. V.) 58P1D12 cellular immune response The mechanism by which sputum cells recognize antigen has been described. The potent peptide epitope determinant vaccine compositions of the invention can elicit a therapeutic or prophylactic immune response in an extremely wide population of the world. In order to understand the value and efficacy of the compositions of the invention to induce cellular immune responses, the present invention provides a brief review of immunologically relevant techniques. The complex of the HL(R) molecule and the peptide antigen acts as a ligand recognized by HL(R) restricted sputum cells (Buus, S. et al, Cell 47: 1071, 1986; Babbitt, BP et al, Nature 3 17: 359, 1985; Townsend , A. and Bodmer, H., Annu. Rev. Immunol. 7:601, 1989 I Germain, RN, Annu. Rev. Immunol. 11:403, 1993). Through the study of single amino acid substituted antigen analogs and the sequencing of endogenously bound natural processing peptides, key residues corresponding to the motifs required for specific binding to HLA antigen molecules have been identified and listed In Table IV (see also, for example, Southwood et al., 142, 769. doc-69-201021828, J. Immunol. 160: 3363, 1998; Rammensee et al, Immunogenetics 41: 178, 1995; Rammensee et al., SYFPEITHI, via the full, ball information Web URL (134.2.96.221/scripts.hlaserver.dll/home.htm) obtained; Sette, A. and Sidney, J.

Curr. Opin. Immunol. 10: 478, 1998; Engelhard, V. H.,

Curr. Opin. Immunol. 6:13,1994; Sette, A_ and Grey, Η. M.,

Curr. Opin. Immunol. 4:79, 1992; Sinigaglia, F' and Hammer, J. Curr. Biol. 6:52, 1994; Ruppert et al., Cell 74: 929-937, 1993; Kondo et al., J. Immunol. 155: 4307-4312, 1995; ❹

Sidney et al, J_ Immunol. 157: 3480-3490, 1996; Sidney et al, Human Immunol. 45: 79-93, 1996; Sette, A. and Sidney, J. Immunogenetics November 1999; ): 201-12, Review) ° In addition, X-ray crystallographic analysis of HLA-peptide complexes has revealed that it is located in the peptide-binding cleft/groove of HLA molecules, and is housed in peptide alleles in an allele-specific pattern. The pocket of the residue; these residues in turn determine the HLA binding capacity of the peptide in which it is present. (See, for example, Madden, DR ® Annu. Rev. Immunol. 13:5 87, 1995; Smith et al, Immunity 4:203, 1996, Fremont et al, Immunity 8:305, 1998; Stern et al., Structure 2: 245, 1994; Jones, EY Curr. Opin. Immunol. 9:75, 1997; Brown, JH et al, Nature 364:33, 1993; Guo, HC et al, Proc. Natl. Acad. Sci. USA 90:8053 , 1993; Guo, H. C. et al., Nature 360: 364, 1992; Silver, ML et al, Nature 360: 367, 1992; Matsumura, M. 142769.doc -70-201021828 et al., Science 257:927 , 1992; Madden et al., Cell 70: 1035, 1992; Fremont, DH et al., Science 257:919, 1992; Saper, MA, Bjorkman, PJA Wiley, DC, J. Mol. Biol. 219:277, 1991) . Thus, the definition of class I and class II allele-specific HLA binding motifs or class I or class II supermotifs allows for the identification of regions within a protein that bind to a particular HLA antigen. Therefore, candidates for epitope-based vaccines have been identified by HLA motif identification methods; such candidates can be further evaluated by HLA peptide binding assays to determine the binding affinity of the epitope and its corresponding HLA molecule and / or combined time. Other confirmatory work can be performed to select among these candidate vaccines epitopes that have better characteristics in terms of population coverage and/or immunogenicity. Cellular immunogenicity can be assessed using a variety of strategies, including: 1) Assess primary T cell cultures from normal individuals (see eg

Wentworth, P. Α· et al, Mol. Immunol. 32:603, 1995;

Celis, E., et al., Proc. Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et al., J. Immunol. 158:1796, 1997; Kawashima, I. et al., Human Immunol. 59 :1, 1998). This procedure involves stimulating peripheral blood lymphocytes (PBL) from a normal subject with a test peptide in vitro in the presence of antigen presenting cells over a period of several weeks. The peptide-specific T cells become activated during this period and can be detected using, for example, a lymphokine factor release assay or a 51Cr release assay involving a peptide-sensitized target cell. 2) Immunization of HLA transgenic mice (see for example Wentworth, PA 142769. doc-71 201023228 et al, J. Immunol. 26:97, 1996; Wentworth, PA et al, Int. Immunol. 8:651, 1996 ; Alexander, J. et al.,

Immunol. 159:4753, 1997). For example, in such methods, peptides in incomplete Freund's adjuvant are administered subcutaneously to HLA transgenic mice. After several weeks of immunization, splenocytes were removed and cultured in vitro for about one week in the presence of test peptides. Peptide-specific T cells are detected using a 5 1 Cr release assay involving, for example, peptide-sensitizing cells and standard stem cells expressing endogenously produced antigens. 3) Verification of recalled T cell responses from immunized individuals who have been effectively vaccinated and/or from chronically ill patients (see, eg, Rehermann, B. et al, J. Exp. Med. 181:1047, 1995; Doolan, DL et al. , Immunity 7:97, 1997; Bertoni, R. et al., J. Clin. Invest. 100:503, 1997; Threlkeld, SC et al., J. Immunol. 159:1648, 1997; Diepolder, Η. M. Man, J. Virol 71: 6011, 1997). Therefore, the recall response is detected by culturing a PBL from an individual exposed to an antigen due to a disease and thus having a "natural" immune response, or culturing a PBL from a patient who has been vaccinated against the antigen. PBL from an individual is cultured in vitro for 1-2 weeks in the presence of test peptide plus antigen presenting cells (APC) to allow activation of "memory" T cells compared to "untreated" T cells. At the end of the incubation period, T cell activity is detected using assays involving 5 1 Cr release, T cell proliferation, or lymphokine release involving peptide sensitization targets. VI.) 5 8P1D12 Transgenic Genes Nucleic acids encoding 58P1D12-related proteins can also be used to produce transgenic genes 142769.doc -72- 201021828 Animals or "gene knockout" animals, which can be used for development and screening. Useful reagents. According to the prior art, genomic DNA encoding 58P1D12 can be cloned using a cDNA encoding 58 piDi2. The transgenic animal can then be produced using the selected genomic sequence, and the transgenic animal 3 has cells expressing the DNA encoding 58P1D12. Methods of producing a transgenic animal, particularly an animal such as a mouse or a rat, are known in the art and are described, for example, in U.S. Patent Nos. 4,736,86, issued on April 12, 1988, and September 1989. U.S. Patent No. 4,870,009 issued on Jun. 26. Specific cells are typically targeted so that the 58P1D12 transgene is combined with a tissue-specific enhancer. The effect of a stronger expression of the DNA encoding 58P1D12 can be examined using a transgenic animal comprising a copy of the transgenic gene encoding Ma 58P1D12. Such animals can be used as test animals which are believed to confer protection against agents such as pathological conditions associated with excessive expression of the DNA. According to this aspect of the invention, the animals are treated with the agent and the reduced incidence of pathological conditions indicates the potential for pathological conditions compared to untreated animals having the transgenic gene. Alternatively, a 58P1D12 "knockout" animal can be constructed using a non-human homolog of 58P1D12, which is homologous between the endogenous gene encoding 5 8 Ρ 1D12 and the altered gene iDNA encoding 58P1D12 introduced into the embryonic cells of the animal. Recombinant with a defective gene or altered gene encoding 58P1D12. For example, genomic DNA encoding 58P1D12 can be cloned according to the prior art using cDNA encoding 58P1D12. One portion of the genomic DNA encoding 58P1D12 can be deleted or replaced with another gene, such as the gene encoding 142769.doc-73. 201021828 for monitoring the selectable marker of integration. The vector typically includes the thousands of assays (5' and 3' ends) of the unmodified flanking DNA (for a description of homologous recombination vectors, see, for example, Thomas and Capecchi, Cell, 51:503 (1987)). The vector is introduced into an embryonic stem cell line (e.g., by electroporation) and a cell in which the introduced DNA has been homologously recombined with the endogenous DNA is selected (see, for example, Li et al., Cell, 69:915 (1992)). The selected cells are then injected into the germ cells of an animal (e.g., mouse or rat) to form aggregate chimeras (see, for example, Bradley, Teratocarcinomas and Embryonic Stem).

Cells: A Practical Approach, edited by E. J. Robertson (IRL,

Oxford, 1987), pp. 113-152). The chimeric embryos can then be implanted into appropriate pseudopregnant female nurturing animals and the embryos formed a "gene knocking" animal in time. Progeny of homologously recombined DNA in germ cells can be identified by standard techniques and can be used to propagate animals, wherein all cells of the animal contain homologously recombined DNA. A knockout animal can be characterized, for example, by its lack of 58P1D12 polymorphism, which is capable of resisting certain pathological conditions or developing pathological conditions. VII.) Method for detecting 58P1D12 Another aspect of the present invention relates to a method for detecting 58P1D12 polynucleic acid and 58P1D12-related protein and a method for identifying 58P1D12 expressing cells. The distribution of the performance of 58P1D12 makes it a diagnostic marker for metastatic disease. Thus, the state of the '58P1D12 gene product provides an S-sense that is suitable for predicting a variety of factors, including advanced disease susceptibility, rate of progression, and/or tumor invasiveness. As detailed herein, the status of the 581>1〇12 gene product in a patient sample can be analyzed by a variety of protocols well known in the art, including 142769.doc • 74· 201021828 including immunohistochemical analysis, various northern inks. Point technology (including in-situ miscellaneous), RT-PCR analysis (eg, for laser capture micro-cut samples), Western blot analysis, and tissue array analysis β are more specific and δ, the present invention provides detection of biological samples (such as serum Characterization of the 58P1D12 polynucleotide in bone, ovary and other tissues, urine, semen, cell preparations and the like. The detectable 58 piD12 polynucleotide includes, for example, the 58P1D12 gene or a fragment thereof, 58ρ1〇ΐ2 mRNA, an alternative splice variant 58Plm2 mRNA, and a recombinant DNA or RNA molecule comprising a 58P1D12 polynucleotide. A variety of methods for amplifying the 58P1D12 polynucleotide and/or detecting the presence of the 5<8>P1D12 polynucleotide are well known in the art and can be used in the practice of this aspect of the invention. In a consistent example, the method of detecting 58P1D12 mRNA in a biological sample comprises: preparing cDNA from a sample by reverse transcription using at least one primer. Using 58P1D12 polynucleotide as a sense of amplification of the 58ρ1〇ΐ2 cDNA therein And an antisense primer to amplify the cDNA prepared thereby; and detecting the presence or absence of φ amplified 58P1D12 cDNA. The sequence of the amplified 58P1D12 cDNA can be determined as needed. In another embodiment, the method for detecting the 58P1D12 gene in a biological sample comprises: firstly isolating genomic DNA from the sample; using 58P1D12 polynucleotide as a sense and antisense primer to amplify the isolated genomic DNA; The presence or absence of the amplified 58P1D12 gene was measured. Many suitable combinations of sense and antisense probes can be designed from the 5<8>P1D12 nucleotide sequence (see, e.g., Figure i) and used for this purpose. The invention also provides assays for detecting the presence of 5 8 卩 1012 protein in tissue or other biological samples such as serum, serum, bone, ovary, urine, cell preparations and the like. Methods for detecting 581 > 1] 〇 12 related proteins are also well known and include, for example, immunoprecipitation, immunohistochemical analysis, Western blot analysis, molecular binding assays, ELISA, ELIFA, and the like. For example, a method of detecting a biological sample for the presence of a 58 piD12-related protein comprises first contacting a sample with a 58p1Ε12 antibody, a 58P1D12 reactive fragment thereof, or a recombinant protein comprising an antigen binding region of a 58P1D12 antibody; and then detecting the sample 581 > 11) The binding of 12 related proteins. Methods for identifying 58P1D12 expressing cells are also within the scope of the invention. In one embodiment, the assay to identify cells expressing the 58P1D12 gene comprises detecting the presence of 58P1D12 mRNA in the cell. Methods for detecting specific mRNA in cells are well known and include, for example, hybridization assays using complementary dna probes (such as in situ hybridization using labeled 58P1D12 nucleic acid probes (riboprobe), northern blots and related techniques) and various Nucleic acid amplification assays (such as RT-PCR using complementary primers specific for 58P1D12, and other amplification-type detection methods such as branched DNA, SISBA, TMA, and the like). Alternatively, the assay to identify cells expressing the 5 8P1D12 gene comprises detecting the presence or absence of a 58P1D12 associated protein in the cell or the presence or absence of a 58P1D12 associated protein secreted by the cell. A variety of methods for detecting proteins are well known in the art and are useful for detecting 58P1D12 related proteins and cells expressing 58P1D12 related proteins. The 58P1D12 performance analysis can also be used as a tool to identify and evaluate agents that modulate the performance of the 58P1D12 gene. For example, 58P1D12 is shown to be significantly up-regulated in ovarian cancer 142769.doc-76·201021828 and is shown in cancers of the tissues listed in Table i. Screening of molecules or biologic agents that inhibit the expression or overexpression of 58P1D12 in cancer cells is of therapeutic value. For example, the agent can be identified by screening using 58P1di2 expression by RT-PCR, nucleic acid hybridization or antibody binding. VIII.) Methods for Monitoring the Status of 58P1D12 Related Genes and Their Products It is known that tumor formation is a multi-step process in which cell growth is gradually dysregulated and cells develop from a normal physiological state to a precancerous state and then develop into a Carcinogenesis® state (see, for example, Alers Et al., Lab Invest. 77(5): 437-438 (1997) and Isaacs et al., Cancer Surv. 23: 19-32 (1995)). In this context 'inspecting evidence of cell growth disorders in biological samples (such as abnormal 58P1D12 expression in cancer) may allow early in the pathological state (such as when cancer has progressed to a stage where treatment options are more limited and/or the prognosis is worse) The abnormal physiological state was detected. In such examinations, for example, the state of 58P1D12 in the desired biological sample can be compared to the state of 58P1D12 in the corresponding normal sample (e.g., a sample of the other body or another body that is not affected by the lesion). A change in the state of 58P1D12 in a biological sample (compared to a normal sample) provides evidence of a disorder in cell growth. In addition to using a biological sample that is not affected by the lesion as a normal sample, a predetermined normal value, such as a predetermined normal amount of mRNA expression, can also be used (see, eg, Grover et al, j Comp. Neurol. December 9, 1996: 3 76 ( 2): 306-14 and US Patent No. 5,83 7,501) to compare the 58卩1〇12 states in the sample. The term "state" in this context is used in accordance with the meaning accepted in the art and refers to the condition or state of the gene and its products. Familiar with this technique 142769.doc -77- 201021828 The surgeon usually uses a variety of parameters to assess the status or status of genes and their products. The parameters of the S-specialization include, but are not limited to, the location of the expressed gene product (including the location of the 58P1D12-expressing cells) and the amount, and the biological activity of the expressed gene product (such as 58P1D12 mRNA, polynucleotide, and polypeptide). The change in the state of 58P1D12 typically involves a change in the position of the 58P1D12 and/or 58P1D12 expressing cells and/or an increase in the expression of 58P1D12 mRNA and/or protein. The 58P1D12 status in the sample can be analyzed by a variety of means well known in the art including, without limitation, immunohistochemical analysis, in situ hybridization, RT-PCR analysis (for laser capture micro-cut samples), Western blot analysis, and Tissue array analysis. Typical protocols for assessing the status of the 58P1D12 gene and gene product can be found, for example, in Ausubel et al., 1995, Current Protocols In Molecular Biology, Unit 2 (Northern Ink Point Method), 4 (Southern Ink Point Method), 15 (immunoblotting) ) and 1 8 (PCR analysis). Thus, the status of 58P1D12 in a biological sample can be assessed by a variety of methods used by those skilled in the art including, but not limited to, genomic Southern analysis (checking for example, perturbations in the 58P1D12 gene); Northern analysis of 58P1D12 mRNA and / or PCR analysis (check for changes in the nucleotide sequence or amount of expression of 58P1D12 mRNA) and Western analysis and / or immunohistochemical analysis (for example, changes in polypeptide sequence, changes in polypeptide localization in the sample, performance of 58P1D12 protein) A change in amount and/or a change in the binding of the 58P1D12 protein to the polypeptide binding partner). The detectable 58P1D12 polynucleotide includes, for example, the 58P1D12 gene or a fragment thereof, 58P1D12 mRNA, an alternative splice variant, 58P1D12 mRNA, and recombinant DNA containing 58P1D12 polynucleotide or 142769.doc •78-201021828 RNA molecule. The performance profile of 58P1D12 makes it a diagnostic marker for local disease and/or metastatic disease and provides information on the growth or carcinogenic potential of biological samples. In particular, the state of 58P1D12 provides information suitable for predicting susceptibility to specific disease stages, development, and/or tumor invasiveness. Methods and assays for determining the status of 58P1D12 and diagnosing cancers exhibiting 58P1D12, such as cancers listed in Table I, are provided. For example, since 58P1D12 mRNA is so highly expressed in nested cancer and other cancers relative to normal, nested tissue, a assay that assesses the amount of 58P1D12 mRNA transcript or protein in a biological sample can be used to diagnose a disorder associated with 58P1D12. The disease, and can provide prognostic information suitable for determining appropriate treatment options. The information provided by the performance status of 58P1D12 includes the presence, stage and location of dysplastic cells, precancerous cells and cancerous cells; prediction of susceptibility to various stages of the disease; and/or determination of tumor invasiveness. In addition, the performance of the knife cloth makes it an imaging agent that can be used as a metastatic disease. Thus, one aspect of the invention wx is a multi-molecular prognosis for examining the state of 58P1D12 in a biological sample, such as from a biological sample of an individual suffering from or suspected of having a pathology (such as cancer) characterized by a disorder of cell growth. And diagnostic methods. The state of 58P1D12 in the biological sample as described above can be examined by a variety of procedures well known in the art. For example, the status of 58P1D12 in a biological sample taken from a particular location in the body can be examined by assessing the presence or absence of 58P1D12 expressing cells (e.g., cells expressing 58 piD12 mRNA or protein) in the sample. For example, winter 142769.doc -79· 201021828 58P1D12 shows that cells present in biological samples (such as lymph nodes) that do not normally contain such cells, this test can provide evidence of cell growth disorders, because in biological samples These changes in the state of 58P1D 12 are often associated with dysregulation of cell growth. In particular, one of the dysregulations of cells is the transfer of cancer cells from a source organ (such as the ovary) to different regions of the body (such as lymph nodes). In this context, evidence of dysregulation of cells is important because, for example, occult lymph node metastasis can be detected in a significant proportion of prostate cancer patients, and such metastasis is associated with known predictors of disease progression (see, for example, Murphy et al. Man, Prostate 42(4): 3 1 5·3 17 (2000); Su et al., Semin. Surg. Oncol. 18(1): 17-28 (2000); and Freeman et al., J Urol 1995 8 Month 154 (2 Pt 1): 474-8). In one aspect, the invention provides a state of the 58P1D12 gene product expressed by cells from an individual suspected of having a disease associated with a disorder of cell growth, such as hyperplasia or cancer, and then the state thus determined A method of monitoring the 58P1D12 gene product in comparison to the status of the 58P1D12 gene product in the corresponding normal sample. The presence of an abnormal 58P1D12 gene product in the test sample relative to the normal sample indicates a dysregulation of cell growth in the individual cell. In another aspect, the invention provides assays suitable for determining the presence or absence of cancer in an individual' such assays comprise detecting a testosterone such as β.

(but not limited to) the presence or absence of evaluable tissue in any of the tissues listed in Table I (including the presence or absence of 58P1D12 mRNA. Since the corresponding normal tissue does not exhibit 5 8ρ丨D丄2 mRNA or 142769.doc *80 - 201021828 Low amounts of 58P1D12 mRNA, so the presence of 58P1D12 in any of these tissues can be used to indicate the presence, presence and/or severity of cancer. In a related embodiment, 58P1D12 is determined at the protein level rather than at the nucleic acid level. For example, the method comprises: determining the amount of 58P1D12 protein expressed by the cells in the test tissue sample and comparing the content thus determined to the 58P1D12 content expressed in the corresponding normal sample. In a consistent example, for example The presence of the <58P1D12 protein is assessed using immunohistochemical methods. The 58P1D12 antibody or binding partner capable of detecting the 58P1D12 protein expression can be used for this purpose in a variety of assay formats well known in the art. In another embodiment, the evaluable organism The state of the 58P1D12 nucleotide and amino acid sequences in the sample to identify the molecular structure Perturbations. These perturbations may include insertions, deletions, substitutions, and the like. Such evaluations are useful because of the observed perturbations in nucleotide and amino acid sequences in a large number of proteins associated with growth dysfunction phenotypes ( See, for example, Marrogi et al, 1999, J. Cutan Pathol 26(8): 369-378. For example, a mutation in the 58P1D12 sequence may indicate the presence or enhancement of a tumor. Therefore, such assays have diagnostic and predictive value, Mutations in 58P1D12 indicate decreased potential function or enhanced tumor growth. A variety of assays for observing perturbations in nucleotide and amino acid sequences are well known in the art. For example, by the description herein Northern, Southern, Western, PCR, and DNA sequencing programs to observe the size and structure of the nucleic acid or amino acid sequence of the 58P1D12 gene product. In addition, observe the perturbations in the nucleotide sequence of nucleocapnic acid and amine 142769.doc -81 - 201021828 Other methods, such as single-strand configuration polymorphism analysis, are well known in the art (see, for example, U.S. Patent No. 5,382,51, issued on September 7, 1999, and 1995! U.S. Patent No. 5,952,170 issued on the 17th. In addition, the thiolation state of the 581>1]〇12 gene in the biological sample can be examined. Gene 5, the abnormal demethylation of the CpG island in the regulatory region and / or hyperthiolation often occurs in immortalized cells and transformed cells, and can cause changes in the expression of a variety of genes. For example, π-type glutathione s_transferase (expressed in the normal prostate, but not The hyperpurification of the promoter of the protein enamel expressed in >9% prostate cancer appears to permanently silence the transcription of this gene and is the most frequently detected genomic alteration in prostate cancer (De Marzo et al. ,

Am. J. Pathol. 155 (6): 1985-1992 (1999)). In addition, this change is present in advanced prostate intraepithelial neoplasia (PIN) cases of at least 70 〇/〇 (Brooks, Cancer Epidemiol, Biomarkers Prev., 1998, 7: 531-536). In another example, the expression of the 'LAGE-I tumor-specific gene (which is not expressed in the normal prostate but expressed in 25_5% of prostate cancer) is induced by deoxyazacytidine in lymphoblastoid cells, This indicates that the tumor ❿ performance is due to demethylation (Lethe et al., 1 this: (&&;11<^76(6).. 903-908 (1998)). Various tests for examining the methylation status of genes have been It is well known in the art. For example, a methylation-sensitive restriction enzyme that does not cleave a sequence containing a thiolated CpG site can be used in a Southern hybridization method to assess the methylation status of a CpG island. (Methylation-specific PCR) allows rapid determination of the thiolation status of all CpG sites present in a CpG island of a given gene. This procedure involves the initial use of sodium bisulfite (which gives 142769.doc-82- 201021828 There is unmethylated cytosine converted to uracil) modified DNA, followed by amplification of methylated DNA relative to unmethylated 〇1 VIII specific primers, including methylation interference can also be found in For example, Current pr〇t〇e〇is ιη Molecular Biology, Unit 12, Freder Ig M Ausubel et al., ed. 1995. Gene amplification is another method for assessing the status of 58P1D12. Using appropriately labeled probes, based on the sequences provided herein, directly, for example, by quantifying the transcription of mRNA Conventional Southern Ink Point Method or Northern Ink Point Method (Thomas, 1980, Pr〇c. Natl. Acad Sci USA, 77:52〇1 5205), point-solving method (DNA analysis) or in situ hybridization method Medium-quantity amplification of genes. Alternatively, antibodies that recognize specific duplexes, including DNA duplexes, RNA duplexes, and 〇>^ RNA hybrids, are used.

A duplex or DNA protein duplex. The antibody is then labeled and assayed, wherein the duplex is bound to the surface such that the duplex is formed on the surface after the presence of an antibody that binds to the duplex is detected. φ can be conveniently used to detect the presence of cancer cells in the biopsy tissue or surrounding blood using, for example, detecting the north of the 58plD12 expression, dot blot, or RT_pCR analysis. The presence of RT-PCR amplifiable 58P1D12 mRNA indicates the presence of cancer. RT-PCR assays are well known in the art. RT_PCR detection assays for tumor cells in peripheral blood are currently evaluated for the diagnosis and control of a variety of human solid tumors. Another aspect of the invention is to assess an individual's susceptibility to developing cancer. In only one example, a method for predicting cancer susceptibility comprises detecting 58P1D12 mRNA or 58P1D12 protein in a tissue sample, the presence of which indicates cancer 142769.doc • 83· 201021828 Susceptibility' wherein the degree of 58P1D12 mRNA expression correlates with the degree of susceptibility. In a specific embodiment, the presence of 58P1D12 in the ovary or other tissue is examined, wherein the presence of 58 pl Di2 in the sample indicates susceptibility to ovarian cancer (or the presence or presence of an ovarian tumor). Similarly, the integrity of the 5<8>P1D12 nucleotide and amino acid sequences in a biological sample can be assessed to identify perturbations in such molecular structures, such as insertions, deletions, substitutions, and the like. One or more perturbations in the 58P1D12 gene product in the sample indicate cancer susceptibility (or the presence or presence of a tumor). The invention also encompasses methods of determining tumor invasiveness. In one embodiment, the method for determining tumor invasiveness comprises: determining the content of 58P1D12 mRNA or 58P1D12 protein expressed by the tumor cells; and determining the content as determined by 58piD12 mRNA or 58P1D12· protein in the corresponding normal tissue. The content was compared. The normal tissue was taken from the same individual or normal tissue reference sample, and the degree of performance of the ten tumor samples relative to the 58P1D12 mRNA or 58ΡΠΜ2 protein in the normal sample indicates the degree of invasiveness. In a specific embodiment, tumor invasiveness is assessed by measuring the extent to which 58 piD12 is expressed in tumor cells, with higher expression indicating greater tumor invasiveness. Another embodiment is to assess the integrity of the 58 pi Di2 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in such molecular structures such as insertions, deletions, substitutions and the like. The presence of one or more perturbations indicates that the tumor is more aggressive. Another embodiment of the present invention relates to a method of observing the development of a malignant disease in an individual over time. In an embodiment, the method of observing an individual's malignant disease over time comprises: determining the amount of protein 142769.doc •84.201021828 58P1D12 mRNA or 581>1〇12 expressed by the cell in the tumor sample; The amount is compared to the amount of 58P1D12 mRNA or 58P1D12 protein expressed in a comparable tissue sample taken from the same individual at different times, wherein the degree of 58P1D12 mRNA or 58P1D12 protein in the tumor sample over time provides information about cancer development. In a specific embodiment, cancer development is assessed by measuring the performance of 58P1D12 over time in tumor cells, wherein increased expression over time indicates cancer progression. In addition, the integrity of the 58P1D12 nucleotide and amino acid sequences in the biological sample can be assessed to identify perturbations in such molecular structures, such as insertions, deletions, substitutions, and the like, in which one or more perturbations are present. Indicates cancer development. The above diagnostic methods can be combined with any of a variety of prognostic and diagnostic protocols known in the art. For example, another embodiment of the present invention relates to a method for observing whether a performance of a 58P1D12 gene and a 58P1D12 gene product (or a disturbance in a 58P1D12 gene and a 58P1D12 gene product) is compatible with a malignant disease phase factor as a diagnosis And the way in which the state of the tissue sample is predicted. Various factors associated with malignant diseases can be utilized, such as the expression of genes associated with malignant diseases and gross cytological observations (see, for example, Bocking et al., 1984, Anal. Quant. Cytol. 6(2): 74-88; Epstein, 1995, Hum. Pathol. 26(2): 223-9; Thorson et al., 1998, Mod. Pathol. 11(6): 543-51; Baisden et al., 1999, Am. J. Surg· Pathol· 23 ( 8): 918-24). For example, a method for observing the performance of the 58P1D12 gene and the 58P1D12 gene product (or the perturbation in the 58p1D12 gene and the 58P1D12 gene product) and another malignant disease-related factor 142769.doc-85-201021828 can be used, Because there is a set of specific factors that are consistent with the disease, it provides key information for diagnosing and speculating the status of tissue samples. Methods for detecting and quantifying the expression of 58p i D丨2 mRNA or protein are described herein, and standard nucleic acid and protein detection and quantification techniques are well known in the art. Standard methods for detecting and quantifying 58P1D12 mRNA include: in situ hybridization using a labeled 58ρ1Ε>12 nucleic acid probe; Northern blotting using a 58P1D12 polynucleotide probe and related techniques; using a specific primer for 58P1D12 Ten RT_PCR* analysis methods; and other amplification type detection methods, such as branch DN A, SISBA, TMA, and the like. In a specific embodiment, semi-quantitative RT-PCR was used to detect and quantify 58 piD12 mRNA expression. Many primers capable of amplifying 58P1D12 can be used for this purpose including, but not limited to, multiple sets of primers specifically described in the private text. In a particular embodiment, a multi-strain or monoclonal antibody that specifically reacts with a wild-type 58P1D12 protein can be used in immunohistochemical assays for tissue testing. IX.) Identifying molecules that interact with 58P1D12梱 The 58P1D12 protein and nucleic acid sequences disclosed herein allow those skilled in the art to identify proteins, small molecules, and other agents that interact with 58P1D12, as well as by 58PID 12 via the art. The pathway of activation of any of the various protocols accepted. For example, one of the so-called interaction intercepting systems (also known as "two-hybrid assay") can be used. In these systems, 'molecular interactions and recombination direct the reporter's expression of transcription factors, thereby characterizing the expression of the reporter gene. Other systems identify intracellular protein-protein interactions by reconstituting eukaryotic transcriptional activators, see, for example, U.S. Patent No. 5,955,28, issued September 21, 1999; July 142, 769.doc -86 - 201021828 20 U.S. Patent No. 5, 925, 523 issued to U.S. Patent No. 5,846,722, issued on Dec. 8, 1998; and U.S. Patent No. 6, s. Algorithms for predicting protein function based on genomes in this technique can also be utilized (see, for example, Marcotte et al., Nature 402: November 4, 1999, 83-86). Alternatively, the peptide library can be screened to identify molecules that interact with the 58P1D12 protein sequence. In such methods, a peptide that binds to 58 pl Di2 is identified by screening a library encoding a set of random amino acids or controlled amino acids. The peptide encoded by the library was expressed as a fusion protein of the phage in vitro protein, and then the phage particle was screened for the 58P1D12 protein. Thus, the appearance of a variety of uses (such as therapeutic, prognostic or diagnostic agents) is thus identified without any prior information about the structure of the desired ligand or receptor molecule. Typical peptide libraries and screening methods that can be used to identify molecules that interact with the 5 8 P1 D12 protein sequence are disclosed, for example, in U.S. Patent No. 5,723,286 issued March 3, 998, and U.S. Patent No. 5,733,731, issued on March 31, 1998. No. Alternatively, the 58P1D12-mediated protein-protein interaction was identified using a cell line expressing 58P1D12. These interactions can be examined using immunoprecipitation techniques (see, e.g., Hamilton B. J. et al. Biochem. Biophys. Res. Commun. 1999, 261: 646-51). The 58P1D12 protein can be immunoprecipitated from the 58P1D12 expressing cell line using an anti-58P1D12 antibody. Alternatively, the His-tagged antibody can be used in a cell strain engineered to express a fusion of 58P1D12 and His-tag (the above vector). The immunosuppressive system can be examined by procedures such as Western blotting, protein 35s_methionine labeling, protein 142769.doc •87-201021828 qualitative microsequencing, silver staining and two-dimensional gel electrophoresis. Protein binding of the complex. Small molecules and ligands that interact with 58p丨D12 can be identified via the relevant examples of such screening assays. For example, small molecules that interfere with protein function can be identified, including interference with 58P1D12-mediated phosphorylation and dephosphorylation, interaction with DNA or RNA molecules (as an indicator of cell cycle regulation), second messenger signaling or A molecule that has the ability to form tumors. Similarly, small molecules that modulate 58P1D12-related ion channels, protein pumps, or cellular communication functions are identified and used to treat patients with cancers that exhibit 58 PLD12 (see © Hille, B, for example, Ionic Channels of Excitable Membranes, 2nd edition, Sinauer Assoc. , Sunderland, MA, 1992). In addition, ligands that modulate 58P1D12 function can be identified based on their ability to bind 58 piD12 and activate reporter constructs. Typical methods are described, for example, in U.S. Patent No. 5,928,868 issued July 27, 999. And including a method for forming a hybridizing ligand, wherein at least one of the ligands is a small molecule. In an illustrative embodiment, 'using a cell engineered to express a 58P1D12 fusion protein and a DNA binding protein will hybridize the ligand/small Molecular and cDNA library transcriptional activation of egg white fusion protein. The cell-inducible gene contains a reporter gene, which is regulated when the first fusion protein and the second fusion protein are close to each other. This event only binds to the hybrid ligand. Hybrid whitening target sites occur only when selecting the target gene. Unknown small molecule or unknown ligand. This method provides a method for identifying a modulator that activates hydrazine to inhibit 58p 丨 D 丨 2. One embodiment of the present invention comprises screening and 58 〇 〇 12 amine 142769. -88- 201021828 Method for Molecular Interaction of Molecular Acid Sequences The method comprises the steps of: contacting a knife group with a 58P1D12 amino acid sequence; and making the molecular group with 581 > 11) 12 amine groups under conditions conducive to interaction Acid sequence interaction; determining the presence or absence of a molecule that interacts with the 58P1D12 amino acid sequence; and then separating the molecule that does not interact with the 58P1D12 amino acid sequence and the molecule that interacts with the 58piD12 amine S under-sequence. In a particular embodiment, the method further comprises purifying, characterizing, and identifying a molecule that interacts with the 58P1D12 amino acid sequence. The identified molecule can be used to modulate the function performed by the 58P1D12. In a preferred embodiment, 58 piE is made. The 12 amino acid sequence is contacted with a peptide library. x.) Therapeutic methods and compositions are identified as being normally present in a limited set of tissues, but also in cancer (such as those listed in Table I) 58P1D12, a protein expressed in cancers, opens up a variety of therapeutic approaches for the treatment of these cancers. It is worth noting that targeted anti-tumor therapies are useful even when targeting Q proteins in normal tissues, even normal living organ tissues. When expressed, the vital organs are organs necessary for life, such as the heart or the colon. Non-living organs are organs that the individual can survive after removal. Examples of non-living organs are the ovaries, breasts, and prostate. For example, Herceptin® is an FDA-approved drug consisting of antibodies that are immunoreactive with proteins known as HER2, HER2/neu and erb-b-2. Herceptin is marketed by Genentech and is a successful anti-tumor agent in business. Herceptin’s sales in 2002 reached nearly $400 million. Herceptin can treat HER2-positive metastatic breast cancer. However, 142769.doc •89· 201021828 However, the performance of HER2 is not limited to these tumors. This protein is also expressed in a variety of normal tissues. In particular, HER2/neu is known to be present in normal kidneys and hearts, and therefore such tissues are present in all human recipients of Herceptin. Latif, Z. et al., B.J.U. International (200 2) 89: 5-9 also demonstrated that HER2/neu is present in normal kidneys. As evidenced in this paper (which assesses whether renal cell carcinoma is a preferred indication for anti-HER2 antibodies (such as Herceptin), both protein and mRNA are formed in benign kidney tissue. It is worth noting that the HER2/neu protein is strongly overexpressed in benign kidney tissue. Although HER2/neu is expressed in life tissues such as the heart and kidney, Herceptin is an FDA-approved and commercially viable drug. The effect of Herceptin on heart tissue (ie, "cardiac poisoning") is only a side effect of treatment. When treating patients with Herceptin alone, significant cardiac toxicity occurs in a very low percentage of patients. To minimize cardiac toxicity, there are stricter access requirements for treatment with HER2/neu. Factors such as the cause of heart disease are assessed before treatment is available. Of particular note is that although the kidney tissue is shown to behave normally and may even perform better than heart tissue, the kidney does not have any detectable Herceptin side effects. In addition, in the normal tissues of many types of HER2, there are few side effects. Only the heart tissue shows any appreciable side effects. Tissues with particularly prominent HER2/neu properties, such as kidneys, have not been the basis of any side effects. In addition, a favorable therapeutic effect of the anti-tumor therapy Erbitux (ImClone) targeting the epidermal growth factor receptor (EGFR) has been found. EGFR is also exhibited in a variety of normal tissues. After the use of anti-EGFR therapeutics, 142769.doc -90· 201021828 in the normal group, there were extremely limited by-products in the weaving. The general side effect of treatment with egfr is severe skin treatment, which is observed in 1% of treatments. Therefore, the target protein is not as good as protein in normal tissues or even normal living tissues! ^ The utility of a therapeutic agent for certain tumors in which the protein is also overexpressed. For example, the performance in living organs is not harmful in and of itself. In addition, organs that are considered to be exempt, such as the anterior parotid gland and ovary, can be removed without affecting life. Finally, some life steamers are not affected by normal organ performance due to immune privileges. Immune privilege 器官 An organ that is isolated from blood by blood-organ barriers and therefore cannot be reached without fatigue therapy. Examples of immune privileged organs are brain and testicles. Therefore, a therapeutic method for inhibiting the activity of the 58P1D12 protein is suitable for patients suffering from cancer exhibiting 58PUM2. These treatments are usually divided into three categories. The first type of method modulates the 58 piD12 energy associated with tumor cell growth, thereby inhibiting or delaying tumor cell growth or inducing tumor cell killing. φ The second type of method encompasses a variety of methods for inhibiting the binding or association of the 58P1D12 protein with its binding partner or with other proteins. A third type of method contains a variety of methods for inhibiting transcription of the 58P1D12 gene or translation of 58P1D12 mRNA. X. A.) Anti-cancer vaccine The present invention provides a cancer vaccine comprising a 58P1D12-related protein or a 58P1D12-related nucleic acid. Considering the performance of 58P1D12, cancer vaccine prevention and/or salty treatment showed that 58P1D12 cancer had minimal or no effect on non-target tissues. Tumor technology for producing a cell-mediated fluid immunoreactivity in vaccines 142769.doc -91- 201021828 Originally known as anti-cancer therapy, it is well known in the art and has been used with nine types of PSMA and rodent PAP immunogens. For use in prostate cancer* (Hodge et al. '1995, Int. J. Cancer 63:231-237; Fong et al., 1997, J. Immunol. 159: 3113-3117). Such methods can be readily carried out by using a 58P1D12-related protein or a nucleic acid molecule encoding 58P1D12 and a recombinant vector capable of expressing and presenting a 58P1D12 immunogen, which typically comprises a plurality of T cell epitopes or antibodies. Those skilled in the art will appreciate that a variety of vaccine systems for delivering immunoreactive antigenic determinants are known in the art (see, for example, ©)

Heryln et al, Ann Med, February 31, 31(1): 66-78; Maruyama et al, Cancer Immunol Immunother, June 2000, 49(3): 123-32). Briefly, the method of generating an immune response (eg, a cell-mediated immune response and/or a humoral immune response) in a mammal comprises the steps of exposing the mammalian immune system to an immunoreactive epitope (eg, present) The 58P1D12 protein or analog thereof shown in Figure 1 is an epitope in a homolog of a homologue" such that the mammal produces an immune response specific for the epitope (eg, produces a specific recognition of the antigen) Antibody). The entire 58P1D12 protein, its immunogenic region or antigenic epitope can be combined and delivered in a variety of ways. Such vaccine compositions may include, for example, lipopeptides (e.g., Vitiello, A. et al., j. Clin. Invest. 95: 341, 1995), encapsulated in poly(dl-lactide-co-glycolide "pLG" ") a composition in a micro-soak (see, for example, Eldridge et al, Molec. Immunol. 28: 287-294, 1991; Alonso et al, Vaccine 12: 299-306, 142769.doc -92-201021828 1994; Jones Et al, Vaccine 13: 675-681, 1995), peptide compositions contained in immunostimulating complexes (ISCOMS) (see, for example, Takahashi et al, Nature 344: 873-875, 1990; Hu et al, Clin Exp Immunol 113:23 5-243, 1998), Multiple Antigen Peptide System (MAP) (see for example Tam, JP, Proc. Natl. Acad. Sci. USA 85: 5409-5413, 1988; Tam, JP, J. Immunol. Methods 196:17-32, 1996), peptides formulated in the form of multivalent peptides, peptides for ballistic delivery systems, typical crystalline peptides, viral delivery vectors (Perkus, Μ. E. et al., in Concepts in vaccine development , Kaufmann, S. Η. E., ed., 379, 1996; Chakrabarti, S. et al., Nature 320: 535, 1986; Hu, SL et al., Nature 3 20:537,1986; Kieny, Μ.-Ρ· et al, AIDS Bio/Technology 4: 790, 1986; Top, FH et al., J. Infect. Dis. 124: 148, 1971; Chanda, PK et al. Virology 175: 535, 1990), from viral sources or synthetic sources (e.g., Kofler, N. et al, J. Immunol. Methods. 192:25, 1996; Eldridge, J. et al., Sem. Hematol. 30 :16,1993 ; Falo, L. D., Jr. et al., Nature Med. 7: 649, 1995), adjuvants (Warren, HS, Vogel, FR and Chedid, LA Annu. Rev. Immunol· 4:369 , 1986; Gupta, RK et al, Vaccine 11: 293, 1993), liposomes (Reddy, R. et al., J. Immunol. 148: 1585, 1992; Rock, Κ·L·, Immunol. Today 17: 131, 1996) or naked cDNA or granule absorption of cDNA (Ulmer, JB et al, Science 259: 1745, 1993;

Robinson, HL, Hunt, L. A., and Webster, RG, Vaccine 11:957, 1993; Shiver, JW, et al., in Concepts in vaccine I42769.doc -93- 201021828 development, Kaufmann, S. Η. E. Edited, p. 423, 1996; Cea.se, KB Sl Berzofsky, JA, Annu. Rev. Immunol. 12: 923, 1994; and Eldridge, JH et al., Sem. Hematol. 30:16, 1993). Toxin targeting delivery techniques, also known as receptor-mediated targeting techniques, such as those of Avant Immunotherapeutics, Inc. (Needham, Massachusetts) can also be used. In patients with 58P1D12-related cancers, this can also be used. The vaccines and antibody compositions of the invention are used in combination with other therapies for cancer (eg, surgery, chemotherapy, drug therapy, radiation therapy, etc.), including with immunological adjuvants (such as IL-2, IL-12, GM-CSF, and It is used in combination with an adjuvant. Cell Vaccine: CTL epitopes can be determined using specific algorithms (e.g., Brown University, BIMAS and SYFPEITHI) that identify peptides within the 58P1D12 protein that bind to the corresponding HLA allele. In a preferred embodiment, the 58P1D12 immunogen comprises one or more amino acid sequences identified using techniques well known in the art, such as those shown in the various tables previously disclosed, or having a class I HLA. a peptide of a 8, 9 or 10 or 11 amino acids as described in Table IV (A), Table IV (D) or Table IV (E) and/or having: > A peptide of 9 amino acids and comprising a class II HLA motif/supermotif (e.g., Table IV (B) or Table IV (C)). As is understood in the art, the class I HLA binding groove is substantially occluded at the ends such that only peptides of a particular size range can fit within the groove and bind, and class I HL A epitopes typically have a length of 8,

9, 10 or 11 amino acids. In contrast, class II HLA binding grooves are substantially open at the ends, so peptides having about 9 or more amino acids can bind to class II 142769.doc -94 - 201021828 HLA molecules. Since there is a difference in the binding groove between the 1 class 1 and the 11 class 11 £8, the class I HLA motif has length specificity, that is, the position 2 of the indole motif is the amino group to carboxyl direction of the peptide. The second amino acid. The amino acid positions in the η-type motif are relative to each other rather than to the entire peptide, i.e., other amino acids may be attached to the amine and/or carboxy terminus of the sequence having the motif. Class II HLA epitopes are typically 9, 1 , 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids or longer than 25 Amino acid. A variety of methods for generating an immune response in a mammal are known in the art (e.g., as a first step in the production of fusion tumors). A method of producing an immune response in a mammal comprises exposing the mammalian immune system to an immunogenic epitope on a protein (e.g., 58P1D12 protein) to produce an immune response. A typical embodiment consists of a method of generating an immune response against 58P1D12 in a host by contacting the host with a sufficient amount of at least one 58P1D12 sputum cell or a cytotoxic sputum cell epitope or analog thereof; and at least After a periodic interval, the host is re-contacted with 5 8Ρ1D12 Β cells or cytotoxic tau cell epitopes or analogs thereof. A specific embodiment consists of a method for producing an immune response against a 58P1D12-related protein or an artificial multi-receptor peptide, the method comprising: administering a 58P1D12 immunogen in the form of a vaccine formulation (eg, 58P1D12 protein or peptide fragment thereof, 58P1D12 fusion protein or the like) Matters, etc.) are administered to humans or to another mammal. Such vaccine formulations typically further contain a suitable adjuvant (see, e.g., U.S. Patent No. 6,146,635) or a universal helper epitope such as the PADRETM peptide (Epimmune Inc" San Diego, CA; see Example 142769.doc-95-201021828 Alexander et al, J. immun〇i. 2000 164(3); 164(3): 1625-1633; Alexander et al, lmmunity 1994 1(9): 751-761; and Alexander et al.' Immunol. Res. 1998 18(2): 79-92). An alternative method comprises generating an immune response against an 58PID 12 immunogen in an individual by the following steps: injecting a DNA molecule comprising a DNA sequence encoding a 58P1D12 immunogen into a muscle of an individual's body in vivo Or the skin, the 〇ΝΑ sequence is operably linked to a regulatory sequence that controls the expression of the DNA sequence; wherein the 〇ΝΑ molecule is sucked by the cell, the DNA sequence is expressed in the cell and an immune response is produced against the immunogen (see, e.g., U.S. Patent No. 5,962,428) In addition, genetic vaccine promoters such as anionic lipids; saponins; lectins; estrogenic compounds; hydroxylated lower alkyl groups; dimethyl hydrazine: and urea. An anti-genotype antibody mimicking 58P1D12 to produce a response against a target antigen. Nucleic Acid Vaccine: The infectious agent composition of the present invention comprises a nucleic acid-mediated form. DNA or RNA encoding the protein of the present invention can be administered to a patient. The genetic immunization method produces a prophylactic or therapeutic humoral immune response and a cellular immune response against a cancer cell expressing 58P1D12. A construct comprising a DNA encoding a 58p 1 d 12-related protein/immunogen and an appropriate regulatory sequence can be directly injected into the muscle of the individual. Or in the skin 'so that the muscle or skin cells absorb the construct and express the encoded 58PLD12 protein/immunogen. Or, the vaccine contains 58ΗΓΗ2 related proteins. The performance of the 58ΡΠΜ2 related protein immunogen leads to prophylactic against cells with 58P1D12 protein Or therapeutic humoral immunity and cellular immunity. A variety of prophylactic 142769.doc-96-201021828 sexual and therapeutic genetic immunization techniques known in the art can be used. Nucleic acid-based delivery is described, for example, in Wolff et al, Science 247:1465 (1990) and US Patent 5,580,859 No. 5,589,466, 5,804,566, 5,739,1 18, 5,736,524, 5,679,647; WO 98/04720. Examples of DNA-based delivery techniques include "naked DNA," promoting (bupivicaine-mediated, polymer-mediated, peptide-mediated) delivery, cationic lipid complexes, and particle-mediated ("gene guns" Or pressure-mediated delivery (see, e.g., U.S. Patent No. 5,922,687). ® The protein of the invention can be expressed via a viral or bacterial vector for therapeutic or prophylactic immunization purposes. A variety of viral gene delivery systems that can be used in the practice of the invention include, but are not limited to, vaccinia virus, fowlpox virus, canarypox virus, adenovirus, influenza virus, gray matter of the polio Inflammatory virus, adeno-associated virus, lentivirus and sindbis virus (see for example Restivo, 1996, Curr. Opin. Immunol. 8:65 8-663; Tsang et al. J. Natl. Cancer Inst. 87 :982-990 (1995)). A non-viral delivery system can also be used by introducing a naked DNA encoding a 58P1D12-related protein into a patient (e.g., intramuscular or intradermal) to induce an anti-tumor response. Vaccinia virus is used, for example, as a vector to express a nucleotide sequence encoding a peptide of the present invention. Upon introduction of the recombinant vaccinia virus into the host, it exhibits a protein immunogenic peptide and thereby induces a host immune response. Vaccinia vectors and methods suitable for use in the immunization protocol are described, for example, in U.S. Patent No. 4,722,848. Another vector is BCG (Bacille Calmette Guerin). The BCG vector is described in Stover et al, Nature 351: 456-460 (1991). Those skilled in the art will be aware of a variety of other vectors suitable for therapeutic administration or immunization of peptides of the invention, such as adenoviral vectors and adeno-associated viral vectors, reverse transcription, as described in 142769.doc-97-201021828. Viral vector, Salmonella typhi vector, detoxified anthrax toxin vector and the like. Thus, gene delivery systems are used to deliver 58P1D12-related nucleic acid molecules. In one embodiment, a full length human 58P1D12 cDNA is used. In another embodiment, a 58P1D12 nucleic acid molecule encoding a specific cytotoxic T lymphocyte (ctl) and/or an antibody epitope is used. From the II vaccine, a variety of ex vivo strategies can also be used to generate an immune response. One method involves the presentation of the 58P1D12 antigen to the patient's immune system using antigen presenting cells (APCs), such as dendritic cells (DCs). Dendritic cells exhibit class I and terpenoid MHC molecules, B7 costimulators, and IL-12, and are therefore highly specialized antigen presenting cells. In prostate cancer, autologous dendritic cells pulsed with peptides of prostate specific membrane antigen (PSMA) are being used in phase I clinical trials to stimulate the immune system of prostate cancer patients (Tjoa et al., 1996, Prostate 28). :65-69; Murphy et al., 1996, Prostate 29:371-380). Thus, in the context of a class I or class II MHC molecule, dendritic cells can be used to present the 58P1D12 peptide to T cells. In one embodiment, the autologous dendritic cells are pulsed with a 58P1D12 peptide capable of binding to a class I and/or class II MHC molecule. In another embodiment, dendritic cells are pulsed with intact 58P1D12 protein. Another embodiment includes the exemplification of the overexpression of the 58P1D12 gene in dendritic cells using a variety of execution vectors known in the art, such as adenovirus (Arthur et al.) People, 1 997,

Cancer Gene Ther· 4:17-25), retrovirus (Henderson et al., 1996, Cancer Res. 56:3763-3770), lentivirus, adeno-associated virus, DNA transfection (Ribas et al., 1997, Cancer Res) 57:2865-2869) or tumor-derived RNA transfection (Ashley et al., 1997, J. Exp. Med. 186:1177-1182). Cells expressing 58P1D12 can also be engineered to express immunomodulators (such as GM-CSF) and can be used as an immunizing agent. X.B.) 58p1D12 5 8P1D12, which is the target of antibody-based therapies, is an attractive target for antibody-based therapeutic strategies. A variety of antibody strategies for stem-to-extracellular and intracellular molecules are known in the art (see, for example, complement-mediated killing and ADCC-mediated killing and the use of intracellular antibodies). Since cancer cells of various lineages express 58P1D12 relative to corresponding normal cells, systemic administration of the 58PiD12 immunoreactive composition is carried out, and the compositions exhibit excellent sensitivity without immunoreactive composition and non-target organ And toxicity, non-specific and/or non-target effects caused by tissue binding. An antibody that specifically reacts with the domain of 58P1D12 can be used in a systemic treatment of a cancer exhibiting 58 piD12 in the form of a conjugate of a toxin or a therapeutic agent or in the form of a naked antibody capable of inhibiting cell proliferation or function. The 58P1D12 antibody can be introduced into a patient such that the antibody binds to 58p1D12 and modulates function, such as interaction with the binding partner, and thereby mediates the destruction of tumor cells and/or inhibits the growth of tumor cells. The mechanisms by which these antibodies exert a therapeutic effect may include complement-mediated cytolysis, antibody-dependent cell cytotoxicity, regulation of physiological functions of 58plD12, inhibition of ligand 142769.doc-99·201021828 binding or signal transduction pathway, regulation of tumor cells Differentiate, alter tumor angiogenic factor distribution and / or apoptosis. Those skilled in the art will appreciate that antibodies can be used to specifically dry up and bind to immunogenic molecules, such as the immunogenic domain of the 58P1D12 sequence shown in Figure 1. Furthermore, it will be appreciated by those skilled in the art that antibodies are typically conjugated to a cellulolytic agent (see, e.g., Slevers et al, Bl. 93: 11 3678. 3 684 (June 1, 1999)). When a cytotoxic agent and/or a therapeutic agent is delivered directly to a cell (such as by binding it to an antibody specific for a molecule expressed by the cell (e.g., 5 8 p 1 d 12 )), the cytotoxic agent is Cells exert a known biological effect (ie, cytotoxicity). A variety of compositions and methods for killing cells using antibody-cytotoxic agent conjugates are known in the art. In the context of cancer, a typical method entails administering a biologically effective amount of a conjugate to a tumor-bearing animal comprising a selected cytotoxic agent and/or therapeutic agent linked to a crude agent (eg, an anti-58P1D12 antibody). The targeting agent binds to a label on the surface of the cell that binds to or localizes to the cell surface (e.g., 58P1D12). A typical embodiment is a method of delivering a cytotoxic agent and/or a therapeutic agent to a cell expressing 58P1D12, which comprises conjugated a cytotoxic agent to an antibody that immunospecifically binds to a 58P1D12 epitope and exposes the cell to an antibody-agent Joint. Another illustrative embodiment is a method of treating an individual suspected of having a metastatic cancer, the method comprising the step of parenterally administering a pharmaceutical composition comprising a therapeutically effective amount of an antibody to a cytotoxic agent, and / or a conjugate of a therapeutic agent. Cancer immunotherapy using an anti-58P1D12 antibody can be performed according to a variety of methods that have been successfully used in the treatment of other types of cancer, including, but not limited to, colon cancer (Arlen et al, 1998, Crit. Rev. Immunol. 1 8:133-138), multiple myeloma (Ozaki et al., 1 997, Blood 90: 3179-3186; Tsunenari et al., 1997, Blood 90: 2437-2444), gastric cancer (Kasprzyk et al., 1992, Cancer Res. 52: 2771-2776), B cell lymphoma (Funakoshi et al., 1996, J. Immunother. Emphasis Tumor Immunol. 19: 93-101), Leukemia (Zhong et al., 1996). , Leuk·Res. 20: 581-589), colorectal fistula cancer (Moun et al, 1994, Cancer Res. 54: 6160-6166; Velders et al, 1995, Cancer Res. 55: 4398-4403) and breast cancer ( Shepard et al., 1991, J. Clin. Immunol. 11:117-127). Some treatments include conjugated naked antibodies to toxins or radioisotopes, such as Y91 or I131, respectively, to an anti-CD20 antibody (eg, ZevalinTM, IDEC Pharmaceuticals Corp. or BexxarTM, Coulter Pharmaceuticals), while other methods include antibody and other treatments. Co-administered, such as Herceptin (trastuzu Mab) and paclitaxel (Genentech, Inc). The antibody can be conjugated to a therapeutic agent. For the treatment of ovarian cancer, for example, 58P1D12 antibody can be administered in combination with radiation, chemotherapy or hormonal resection. The antibody can also be conjugated to a toxin, such as a conjugated to calicheamicin (eg, MylotargTM 'Wyeth-Ayerst, Madison, NJ, a recombinant humanized IgG4 κ antibody conjugated to the anti-tumor antibiotic echinomycin) or with meiden Alcohol bonding (e.g., a taxane-based tumor activating prodrug TAP platform, ImmunoGen, Cambridge, MA; see also, e.g., U.S. Patent 5,416,064) or conjugated with Auristatin E (Nat 142769.doc-101 - 201021828

Biotechnol· July 2003; 21 (7): 778_84. (SeaUle

Genetics)) 5 Although 5 8 P1D12 antibody therapy can be used in all stages of cancer, antibody therapy is especially useful for advanced or metastatic cancer. Treatment with the antibody therapy of the invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, the antibody therapy of the invention may be combined with chemotherapy or radiation therapy for patients who have not been treated with chemotherapy. In addition, antibody therapy allows the use of low doses of concomitant chemotherapy' especially for patients who are not well toxic to chemotherapeutic agents. Fan et al. (Cancer Res 53: 4637-4642, 1993), Prewett (International J. 〇f 〇nco. 9: 217-224 1996) and Hancock et al. (Cancer Res. 51:4575-4580, 1991) Description Use of various antibodies with chemotherapeutic agents. Although 58P1D12 antibody therapy can be used in all stages of cancer, antibody therapy may be particularly useful for advanced or metastatic cancer. The treatment with the anti-physiological treatment of the present invention is suitable for patients who have received one or more rounds of chemotherapy. Alternatively, the antibody therapy of the invention may be combined with chemotherapy or radiation therapy for patients who have not been treated with chemotherapy. In addition, antibody therapy allows the use of low doses of concomitant chemotherapy, especially in patients who are not well toxic to chemotherapeutic agents. The presence and extent of 58 P1D12 expression in cancer patients can be assessed, preferably using immunohistochemical evaluation of tumor tissue, quantification of 58P1D12 imaging, or other techniques that reliably demonstrate the presence and extent of 58P1D12 performance. Immunohistochemical analysis of tumor biopsy or surgical samples is preferred for this purpose. Immunohistochemical analysis methods for tumor tissues are well known in the art. 142769.doc -102- 201021828 Anti-58P1D12 monoclonal antibodies for the treatment of nested and other cancers include antibodies that elicit a strong immune response against tumors or antibodies that are directly cytotoxic. In view of this, the anti-58P1D12 monoclonal antibody (MAb) can induce tumor cell lysis by complement-mediated or antibody-dependent cell cytotoxicity (ADCC) mechanism. Both of these mechanisms are required for effector cell Fe on the complement protein. The integrity of the immunoglobulin molecule interacting with the body site is in addition to the '58P1D12 _ MAb', which has a direct biological effect on tumor growth, and can be used to treat cancers exhibiting 58 piD12. The direct mechanism of cytotoxic MAb includes: inhibition of cell growth; regulation of cell differentiation; regulation of tumors, production factor distribution; and induction of apoptosis. The mechanism by which a particular anti-58P1D12 MAb exerts an anti-tumor effect can be assessed using a variety of in vitro assays (such as AdCc, ADMMC, complement-mediated cytolysis, etc.) commonly known in the art to assess cell death. In some patients, the use of murine or other non-human monoclonal antibodies or human/mouse chimeric MAbs can induce a moderate immune response against non-human antibodies. This can result in antibody clearance and reduced efficacy. In the most severe cases, this immune response results in the formation of large amounts of immune complexes that can potentially cause kidney failure. Thus, a preferred monoclonal antibody for use in the methods of treatment of the present invention is a single human or humanized and specifically binds to the target 58P1D12 antigen with high affinity in a patient, but exhibits low antigenicity or does not exhibit antigenicity. Strain antibody. Therapeutic methods of the invention encompass the administration of a combination or mixture of a single anti-58P1D12 MAb and a different MAb (i.e., 58P1D12 MAb or a Mab that binds to another protein). Such MAb mixtures may have certain advantages due to the inclusion of MAbs that target different antigens, the use of different effector mechanisms, or the combination of direct cytotoxic MAbs with MAbs that are dependent on immune effector functions. These combined forms of MAb can exhibit a synergistic therapeutic effect. In addition, 58p1D12 MAb may be administered in conjunction with other therapeutic modalities including, but not limited to, various chemotherapeutic agents and biological agents, androgen blockers, immunomodulators (eg, IL-2, GM-CSF). , surgery or radiation. The 58 piDi2 MAb can be administered in its "naked" or unconjugated form, or can have a therapeutic agent associated therewith. The 58P1D12 Mab formulation can be administered via any route capable of delivering antibodies to tumor cells. The route of administration includes, but is not limited to, intravenous, intraperitoneal, intramuscular, intratumoral intradermal and the like. Treatment typically involves repeated administration of a 58P1D12 Mab formulation via an injectable route of administration, such as, but not limited to, 〇1, 〇2, 0.3, 0.4, 0·5, 〇·6, 〇 ·7, 〇·8, 〇9, i, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or 25 mg/kg body weight. In general, 10 per week The dose of -1000 mg MAbi is effective and well tolerated. Based on the clinical experience of treating Hereditary breast cancer with Hercynian and Trastuzumb (Trastuz_b), about 4 mg/kg of MAb preparation The intravenous initial loading dose of the patient's body weight, followed by an intravenous dose of about 2 mg/kg per week, represents an acceptable dosage regimen. The initial loading dose is preferably administered as an infusion of 90 minutes or longer than 90 minutes. Maintain the dose of money for a minute or longer than the μ minute (four), and the _ condition is that the initial dose is well received. As is familiar to those skilled in the art, in the case of (4), a variety of 142769.doc -104- 201021828 Factors can affect the ideal dosing regimen. These factors include, for example, the MAb used. Affinity and half-life; degree of 58P1D12 expression in patients; degree of circulating 58P1D12 antigen; desired steady-state antibody concentration; frequency of treatment; and effects of combination of chemotherapeutic or other agents with the therapeutic methods of the invention' and specific patients Health status. The amount of 58P1D12 in a given sample of the patient (eg, the amount of circulating 58P1D12 antigen and/or 58P1D12 expressing cells) should be assessed as needed to help determine the most effective regimen, etc. Use for monitoring purposes throughout the therapy and with other parameters (eg, urine cytology and/or immune cell assay (ImmunoCyt) levels in bladder cancer treatment, or similarly, serum PSA levels in prostate cancer treatment) The evaluation combination is used to determine the success of the treatment. The anti-genetic anti-58P1D12 antibody can also be used as a vaccine against the cell-induced immune response of the 58P1Di2-related protein for anti-cancer therapy. In particular, the production of anti-genetic antibodies has been Well known in the art; this method can be readily adapted to produce analog 58P1D12 related proteins The original anti-genetic anti-58P1D12 antibody (see, for example, Wagner et al. 1997, Hybridoma 16: 33-40; Foon et al, 1995, J. Clin. Invest. 96: 334-342; Herlyn et al, 1996, Cancer Immunol. Immunother. 43:65_76). Such anti-genetic antibodies can be used in cancer vaccine strategies. It is an object of the present invention to provide a 58P1D12 MAb which inhibits or delays the growth of tumor cells expressing 58P1D12. Another object of the present invention is to provide for the use of such 58P1D12 MAbs and, in particular, the 58P1D12 MAbs in combination with their I42769.doc-105-201021828 other drug or immunological therapies to inhibit angiogenesis and other biological functions and thereby cause mammals ( Methods for slowing tumor growth in better humans' These other drugs or immunological therapies include (but are not limited to): Avastin® (bevacizumab), Sutent® (sunitinib) Malate)), Nexavar® (Sorafinib tosylate), Taxotere® (docetaxel), Sirolimus® (rapamycin or its analogues), Paraplatin® (carobplatin), interleukin-2 (also known as Proleukin®, IL-2 or Aldesleukin) or interferon alpha (interferon-a-2a or interferon-a-2b) Other drugs known in the art to treat cancer. In one embodiment, there is a synergistic effect when treating tumors, including human tumors, with a 58P1D12 antibody in combination with a chemotherapeutic agent or radiation or a combination thereof. In other words, when combined with a chemotherapeutic agent or radiation or a combination thereof, the inhibition of tumor growth by the 58P1D12 antibody is more than expected. Synergism can be presented as & e.g., combination therapy has greater inhibition of tumor growth than the therapeutic expectation of only the 58P1D12 antibody or compared to the additive effect of the 58P1D12 antibody and chemotherapeutic agent or radiation therapy. The synergistic effect is preferably demonstrated by cancer remission. The treatment with naked 581 > 1〇12 antibody or the combination of 58P1D12 antibody and chemotherapeutic agent or radiation is not alleviated. A method of inhibiting tumor cell growth using a 58P1D12 antibody and chemotherapy or radiation, or a combination of both, is included before, during or after the initiation of chemotherapy or radiation therapy, and any combination thereof (ie, at the beginning of chemotherapy and / 142769.doc - 106-201021828 or before and during radiotherapy; before and after; during and after; or before, during and after) 58P1D12 antibody. For example, the 58P1D12 antibody is typically administered between 1 and 60 days, preferably between 3 and 40 days, and more preferably between 5 and 12 days prior to initiation of radiation therapy and/or chemotherapy. However, depending on the treatment regimen and the needs of the particular patient, the method can be performed in a manner that provides the most effective treatment and ultimately extends the life of the patient. Administration of chemotherapeutic agents can be accomplished in a variety of ways, including systemic administration via parenteral and enteral routes. In one embodiment, the 58P1D12 antibody and the ® chemotherapeutic agent are administered as separate molecules. In another embodiment, the 58P1D12 antibody is linked to the chemotherapeutic agent by, for example, bonding. (See the example entitled Human Clinical Trial for Treatment and Diagnosis of Human Carcinoma via the Use of 58P1D12 MAb). Specific examples of chemotherapeutic agents or chemotherapy include shunming, dacarbazine (DTIC), dactinomycin, mechlorethamine (nitrogen mustard), and chain stellate (streptozocin), cyclopamine, carmustine (BCNU), lomustine (CCNU), doxorubicin (adriamycin), daunorubicin ), procarbazine, mitomycin, cytarabine, etoposide, methotrexate, 5-fluorourine ), vinblastine, vincristine, bleomycin, paclitaxel (taxol), docetaxel (taxotere) , aldileukin, aspartate, aspartate, busulfan 142769.doc • 107- 201021828 (busulfan), kaming, cladribine, dakarta, floxuridine, darabine (fludarabine), transurea urea, isocyclic phosphoniumamine, dry Prime alpha, leuprolide, megestrol, melphalan, mereaptopurine, plicamycin, mitotane, culture Pegaspargase, pentostatin, pipobroman, pucamycin, streptozin, tamoxifen, teniposide, sputum S (testolactone), thioguanine thioguanine, thiotepa, uracil mustard, vinorelbine, chlorambucil, The radioactive source used in combination with paclitaxel and its combination 0 and 58P1D12 Mab can be external or internal to the patient being treated. When the source of radiation is outside the patient, this treatment is called extracorporeal radiation therapy (EBRT). When the source is inside the patient, this treatment is called brachytherapy (BT). The above treatment regimens may be further combined with other cancer therapeutics and/or regimens, such as other chemotherapy, cancer vaccines, signal transduction inhibitors, agents useful for treating abnormal cell growth or cancer, Antibodies (such as the anti-CTLA-4 antibodies described in WO/2005/092380 (Pfizer)) or other ligands that inhibit tumor growth by binding to IGF-IR and cytokines. The above chemotherapeutic agents can be used when the mammal is subjected to other chemotherapy. In addition, growth factor inhibitors, biological response modifiers, anti- 142769.doc-108-201021828 hormone therapy, selective estrogen receptor modulator (SERM), angiogenesis inhibitors, and antiandrogens can be used. For example, an anti-hormone such as an anti-estrogen such as Nolvadex (tamoxifen) or an anti-androgen such as Casodex (f-cyano-3-(4-fluorophenylsulfonyl)-2- can be used. Hydroxy-2-methyl-3'-(trifluoromethyl)propananilide). In certain embodiments of the invention, the above methods can be combined with a cancer vaccine. Useful vaccines can be, without limitation, including cancer-associated antigens (eg, BAGE, carcinoembryonic antigen (CEA), EBV, GAGE, gplOO (especially including ® gpl00:209-217 and gpl00:280-288), HBV, HER-2 /neu, HPV, HCV, MAGE, mammaglobin, MART·1/Melan-A, Mucin-1, NY-ESO-1, Protease-3, PS A, RAGE, TRP -1, TRP-2, cool aminase (eg, citrate: 368-376), WT-1), vaccines based on GM-CSF DNA and cells, dendritic cell vaccine, recombination Virus (eg bovine cancer virus) vaccine and heat shock protein (HSP) vaccine. Useful vaccines also include tumor vaccines, such as those formed by melanoma cells, and may be autologous or allogeneic. The vaccine can be based, for example, on peptides, DNA or cells. The plurality of agents may be combined so that a combination comprising, inter alia, gplOO peptide, tyrosinase and MART-1 can be administered together with the antibody. The vaccine can be administered before or after stem cell transplantation, and when chemotherapy is part of the therapy, the vaccine can be administered prior to chemotherapy. In certain embodiments, the antibodies of the invention may also be administered prior to chemotherapy. Vaccines can also be administered after stem cell transplantation and, in certain embodiments, with antibodies. 142769.doc -109- 201021828 The above treatments may also be used in combination with signal transduction inhibitors, such as agents that inhibit EGFR (epidermal growth factor receptor) responses, such as EGFR antibodies, EGF antibodies, and EGFR inhibitor molecules; VEGF (vascular endothelium) Growth factor) inhibitors, such as VEGF receptors and molecules that inhibit VEGF; and erbB2 receptor inhibitors, such as organic molecules or antibodies that bind to the erbB2 receptor. EGFR inhibitors are described, for example, in WO 95/19970 (published July 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO 98/02434 (published Jan. 22, 1998), and U.S. patents. No. 5,747,498 (issued May 5, 1998), and the same may be used in the present invention as described herein. © EGFR inhibitors include, but are not limited to, monoclonal antibodies ERBITUX (ImClone Systems Incorporated, New York, NY) and VECTIBIX (Amgen, Thousand Oaks, Calif.), compound ZD-1839 (AstraZeneca) 'BIBX-1382 (Boehringer) Ingelheim) 'MDX-447 (Medarex Inc., Annandale, New. Jersey) and OLX-103 (Merck & Co., Whitehouse Station, NJ) 'VRCTC-310 (Ventech Research) and EGF fusion toxin (Seragen Inc., Hopkinton, Mass.). These and other EGFR inhibitors can be used in the present invention. VEGF inhibitors (e.g., SU-5416 and SU-6668 (Sugen Inc., South San Francisco, Calif.)) can also be used in combination with antibodies. VEGF inhibitors are described, for example, in WO 99/24440 (published May 20, 1999), PCT International Application PCT/IB99/00797 (filed May 3, 1999), WO 95/21 613 (August 17, 1995) Japanese Open), WO 99/61422 (published December 2, 1999), US Patent No. 5,834,504 (November 10, 1998, 142, 769. doc • 110-201021828), WO 98/503 56 (1998) US Patent No. 5,883,113 (issued March 16, 1999), US Patent No. 5,886,020 (1" issued on March 23, 9), US Patent No. 5,792,783 (August 11, 1998) Promulgated), WO 99/10349 (published March 4, 1999), WO 97/32856 (published on September 12, 1997), WO 97/22596 (published on June 26, 1997), WO 98/54093 ( Published on December 3, 1998, WO 98/02438 (published on January 22, 1998), WO 99/16755 (published on April 8, 1999), and WO 98/02437 (published on January 22, 1998) in. Other examples of certain specific VEGF inhibitors useful in the present invention are IM862 (Cytran Inc., Kirkland, Wash.); IMC-1C11 Imclone antibody and angiozyme (aniozyme) (from species Ribozyme (Boulder, Colo.) ) Synthetic ribozyme with Chiron (Emeryville, Calif.).

ErbB2 receptor inhibitors such as GW-282974 (Glaxo Wellcome) and monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc., The Woodlands, Tex.) and 2B-1 (Chiron) can additionally be specified, for example, in the following patents. Antibody combinations: WO 98/02434 (published January 22, 1998), WO 99/35146 (published July 15, 1999), WO 99/35132 (published July 15, 1999), WO 98/02437 (published on January 22, 1998), WO 97/13 760 (published on April 17, 1997), WO 95/19970 (published on July 27, 1995), and US Patent No. 5,587,458 (December 24, 1996)曰 promulgated) and US Patent No. 5,877,305 (issued March 2, 1999). ErbB2 receptor inhibitors suitable for use in the present invention are also described in EP1029853 (published on Aug. 23, 2000) and WO 00/44728 (published on Aug. 3, 2000). The erbB2 receptor inhibitor compounds and substances described in the above-mentioned PCT application, U.S. Patent, and U.S. Patent No. 142, 769. Therapeutic regimens of the invention may also be combined with antibodies or other ligands that inhibit tumor growth by binding to IGF-IR (insulin-like growth factor 1 receptor). The specific anti-IGF-1R antibodies that can be used in the present invention include those described in PCT Application No. PCT/US01/51113, filed on Dec. 20, 2001, and the disclosure of WO 02/053596. Therapeutic regimens described herein can be combined with anti-angiogenic agents (such as MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase). II) Inhibitors) are combined and can be used in conjunction with antibodies in the methods of the invention. Examples of useful COX-II inhibitors include CELEBREX (celecoxib), valdecoxib, and rofecoxib. X.C.) 58P1D12 as a target for cellular immune responses Vaccines containing one or more immunogenic effective amounts of HL A binding peptides as described herein and methods of preparing such vaccines are other embodiments of the invention. Furthermore, the vaccine of the invention encompasses a composition of one or more of the claimed peptides. The peptide can be present in the vaccine alone. Alternatively, the peptide may comprise a homopolymer form of multiple copies of the same peptide or be present as a heteropolymer of various peptides. The advantage of the polymer is that the immune response is enhanced; and, if different peptide epitope-constituting polymers are used, it is additionally capable of inducing antibodies that react with different antigenic determinants of the pathogenic organism or tumor-related peptide to which the immune response is directed and/or Or CTL. The composition may be a naturally occurring region of the antigen or may be prepared, for example, by the method of group 142769.doc • 112·201021828 or by chemical synthesis. Vectors useful in the vaccines of the invention are well known in the art and include, for example, thyroglobulin, albumin (such as human serum albumin), tetanus toxoid, polyamino acids (such as poly-L-lysine, Poly-L- faceamine), influenza, hepatitis B virus core protein and the like. The vaccine may contain physiologically tolerable (i.e., acceptable) dilutions such as water or saline, preferably phosphate buffered saline. Vaccines usually also include adjuvants. Adjuvants such as incomplete * adjuvant, bismuth phosphate, chlorinated or citric acid are examples of materials well known in the art. Furthermore, as disclosed herein, by binding the peptide of the present invention to a lipid such as trisylsulfonyl-s-glycerylcysteine-mercaptosylamine-serine (PASS) Sensitive CTL reaction. In addition, adjuvants such as synthetic nucleotides containing cytosine-phosphorothioate-guanine (CpG) have been found to increase the CTL response by a factor of 1 to ι〇0 (see, for example, Davila &

Celis, J. Immunol. 165:539-547 (2000)). After immunization of the peptide composition of the present invention by injection, spray, oral, transdermal, transmucosal, intrapleural, intraorbital or other appropriate route, the host's immune system produces a large amount of CTL specific for the desired antigen and / or HTL to respond to the vaccine. Thus, the host becomes at least partially immune to cells that later form or overexpress the 58P1D12 antigen, or at least a certain therapeutic benefit when the antigen is associated with the tumor. In some embodiments, it is desirable to combine a class I peptide component with a component that induces or facilitates a reaction against a target antigen neutralizing antibody and/or helper T cell. A preferred embodiment of the composition comprises the steroid and steroid epitopes of the invention. An alternative embodiment of the composition comprises a class I and/or class II 142769.doc • 113. 201021828 epitope of the invention, and a cross-reactive HTL epitope such as PADREtm (Epimmune, San Dieg〇, CA) Molecules (for example, as described in U.S. Patent No. 5,736,142). The vaccine of the present invention may also include antigen presenting cells (Apc) such as dendritic cells (DC) as a carrier for presenting the peptide of the present invention. The vaccine composition can be produced in vitro after the dendritic cells have moved and harvested, wherein the loading of the dendritic cells is performed in vitro. For example, dendritic cells are transfected with, for example, a minigene of the invention or pulsed with a peptide. Dendritic cells can then be administered to the patient to induce an immune response in vivo. Vaccine compositions (DNA-based or peptide-based) in vivo administration can also be combined with dendritic cell movement, wherein the dendritic cells are carried in vivo. Fortunately, when an array of epitopes is selected for inclusion in a multi-antigen thiol composition for use in a vaccine, or a discrete antigen to be incorporated into a vaccine and/or to be encoded by a nucleic acid (such as a minigene) is selected. At the time of the base, the following principles are used. The following principles are better weighed to make choices. The plurality of epitopes to be incorporated into a given vaccine composition can be, but are not necessarily, the sequence of the natural antigen from which the antigen/thiol is derived. 1) Select the antigenic determinant that mimics the observed immune response associated with tumor elimination after administration. For class 1 11]^8, this includes 3_4 epitopes from at least one tumor-associated 彳/u original (TAA). A similar basic principle is used for the η-type hla; again, 3_4 epitopes are selected from at least one TAA (see, for example, Rosenberg et al. ' science 278: 1447-1450). An epitope from a TAA can be used in combination with an epitope derived from one or more other ΤΑA to prepare a vaccine that is directed to a tumor having a different pattern of expression of TAA that is frequently subjected to 142769.doc • 114-201021828. 2) Select the antigenic determinant associated with the necessary binding affinity for immunogenicity: for class I HLA, IC5Q is 500 η Μ or less than 500 η Μ, usually 200 η Μ or less than 200 η Μ; and for steroid HLA, The redundancy is 1〇〇〇ηΜ or less than 1〇〇〇ηΜ. 3.) Select sufficient peptide arrays with supermotifs or sufficient peptide arrays with allele-specific motifs to achieve broad population coverage. For example, preferably ❹ has a population coverage of at least 80%. The breadth or redundancy of population coverage can be assessed using Monte Carlo anaiysis (a statistical assessment known in the art). 4) When an epitope is selected from a cancer-associated antigen, it is generally useful to select an analog since the patient can form a tolerance to a natural epitope. 5)) A particularly relevant epitope is called a "nested epitope". Nested epitopes are present where at least two epitopes in a given peptide sequence overlap. The nested peptide sequence may comprise a B cell, a class 1 <RTIgt; When a nested epitope is provided, it is generally intended to provide a maximum number of epitopes per sequence. Therefore, the monomorphism avoids providing a peptide longer than the amino terminus of the amino terminal epitope of the peptide and the carboxyl terminus of the carboxyl terminal epitope. When multiple epitope sequences (such as sequences comprising nested epitopes) are provided, screening sequences are often important to ensure that the sequence does not have pathological or other deleterious biological properties. 6.) If a multi-antigenic protein is produced or a minigene is produced, the objective is to produce a minimal peptide covering the desired epitope, at 142769.doc -115-201021828. This principle is similar to the principle used when selecting a peptide comprising a nested epitope. However, there is a trade-off between the need to minimize size for artificial multi-receptor peptides and any intervening sequences between antigenic determinants in the integration of multiple epitope proteins. For example, a spacer amino acid residue can be introduced to avoid a zygosing epitope (an epitope determined by the immune system, in the absence of a target antigen, and only by artificially merging the epitope), or Promotes division between epitopes and thereby enhances epitope presentation. A zygosity epitope is generally avoided because the recipient can produce an immune response against a non-native ◎ epitope. As a "binding antigen" determinant epitope, it is particularly worthy of attention. A dominant antigenic determinant can cause a positive response, thereby attenuating or inhibiting an immune response to other epitopes. 7.) If the sequence of the plurality of variants exists in the same target protein, the potential peptide epitope can also be selected based on its conservation. For example, a conservative standard can be defined as a complete sequence of a class I HLA-binding peptide or a complete 9-brace core of a class-binding peptide that can be conserved in a specified percentage of the sequence specific for the specific protein antigen©. X - C. 1, a small gene vaccine can utilize a variety of different methods that allow simultaneous delivery of multiple epitopes. A nucleic acid encoding a peptide of the present invention is a particularly useful embodiment of one of the inventions. Preferably selected according to the criteria set forth in the previous section. A preferred method of administering a nucleic acid encoding a peptide of the invention uses a minigene 142769.doc-116-encoding a peptide comprising one or more epitopes of the invention. 201021828 Constructs. The use of multiplex epitope minigenes is described below and in the following literature: Ishioka et al, J. Immunol. 162: 3915-3925, 1999; An, L. and Whitton, JL, J. Virol. : 2292, 1997; Thomson, SA et al, J. Immunol. 157:822, 1996; Whitton, JL et al, J. Virol 67:348, 1993; Hanke, R. et al., Vaccine 16:426, 1998 For example, edit The code is derived from the polymorphism DNA and plastid of 58P1D12 with a superbasic and/or motif epitope (PADRETM universal helper T cell® epitope or multiple htl epitopes from 58P1D12) The network displacement signal sequence can be engineered. Vaccines can also contain epitopes obtained from other TAAs. The immunogenicity of multiple epitope determinants can be demonstrated in transgenic mice to assess the antigens tested. The size of the Ctl-induced reaction. In addition, the immunogenicity of the DNA-encoding epitope in vivo can be correlated with the in vitro reaction of the specific CTL line against the target cells transfected with the human plastid. Therefore, Such experiments may demonstrate 1): a minigene is used to generate a CTL response; and 2) an induced CTL recognizes a cell that exhibits an encoded epitope. For example, 'to generate a selected antigen encoding for expression in a human cell The DNA sequence of the determinant (small gene) can reverse the amino acid sequence of the epitope. The human codon usage table can be used to guide the codon usage of each amino acid. The DNA sequence of the epitope epitope can be directly ligated to produce a contiguous polypeptide sequence at turn. To optimize performance and/or immunogenicity, other elements can be incorporated into the minigene design. Reversible translation and inclusion in 142769. Examples of amino acid sequences in the gene sequence of doc-117-201021828 include: HLA epitopes, class II HLA epitopes, antibody epitopes, ubiquitination signal sequences and/or endoplasmic reticulum targeting signals . In addition, HLA presentation of CTL and HTL epitopes can be improved by including a synthetic flanking sequence (eg, polyalanine) adjacent to a CTL or HTL epitope or a naturally occurring flanking sequence; Such larger peptides are within the scope of the invention. The minigene sequence can be converted to DNA by assembling the positive and negative oligonucleotides encoding the minigene. Overlapping nucleotides (30-1 碱基 base length) can be synthesized, squashed, purified, and affixed under appropriate conditions using well-known techniques. The ends of the oligonucleotides can be joined using, for example, T4 DNA ligase. This synthetic minigene encoding the epitope polypeptide can then be cloned into the desired vector. Preferably, the vector comprises standard regulatory sequences well known to those skilled in the art to ensure performance in the target cell. Several vector elements are required: a promoter with a downstream selection site for minigene insertion; a polyadenylation signal that effectively terminates transcription; an E. coli origin of replication; and an E. coli selectable marker (eg, ampicillin) Or kanamycin resistance). A variety of promoters, such as the human cell giant virus (hCMV) promoter, can be used for this purpose. For other suitable promoter sequences, see, for example, U.S. Patent Nos. 5,580,859 and 5,589,466. Additional vector modifications may be required to optimize minigene expression and immunogenicity. In some cases 'effective gene expression requires an intron and one or more synthetic or naturally occurring introns can be incorporated into the transcribed region of the minigene. 142769.doc -118- 201021828 It is also possible to test 3: the mRNA stabilization sequence and the replication sequence into human mammalian cells to enhance minigene expression. The expression vector, selected, is placed downstream of the polymorphic cleavage site linker (P〇lylinker) region of the minigene. Transforming this plastid into an appropriate E. coli strain and preparing the DNA using standard techniques>> using restriction mapping and DNA sequence analysis to demonstrate the orientation of the minigene and the DNA sequence and the vector included in the vector All other components. Bacterial cells containing appropriate plastids can be stored as a master cell bank and a working cell bank. Furthermore, the immunostimulatory sequence (158 or CpG) appears to play a role in the immunogenicity of DNA vaccines. In addition to the minigene coding sequences, such sequences may be included in the vector to enhance immunogenicity, if desired. In some embodiments, a bicistronic expression vector can be used which permits the production of a minigene-encoded epitope and a second protein (incorporated for enhancing or reducing immunogenicity). Examples of proteins or polypeptides that advantageously enhance the immune response if co-expressed include cytokines (eg, IL-2, IL-12, GM-CSF), cytokine-inducing molecules (eg, LelF), costimulatory molecules, or against HTL Reactive pan-DR binding protein (PADRETM, Epimmune, San Diego, CA). The helper (HTL) epitope can be linked to the intracellular targeting signal and separately from the expressed CTL epitope; this allows the HTL epitope to be directed to a cellular metabolic region different from the cellular metabolic region of the CTL epitope. This can more efficiently promote the entry of the HTL epitope into the class II HLA pathway, as needed, thereby improving HTL induction. In certain diseases, it may be advantageous to specifically reduce the immune response by co-expression of immunosuppressive molecules (e.g., TGF-[beta] in comparison to HTL or CTL induction. 142769.doc • 119· 201021828 The therapeutic amount of plastid DNA can be prepared, for example, by fermentation in E. coli and subsequent purification. The growth medium was inoculated using an aliquot from the working cell bank and grown to saturation in a shake flask or bioreactor according to the skill of the recipe. The plastid DNA can be purified using standard biological separation techniques such as solid phase anion exchange resins supplied by QIAGEN, Inc. (Valencia, California). When desired, the supercoiled DNA can be separated from the open-loop form and the linear form using gel electrophoresis or other methods. Purified plastid DNA can be prepared for injection using a variety of formulations. The simplest of these is lyophilized DNA reconstituted in sterile phosphate buffered saline (PBS). This method (referred to as "naked DNA") is currently used in intramuscular (IM) administration in clinical trials. In order to maximize the immunotherapeutic effect of minigene DNA vaccines, alternative methods of purifying purified plastid DNA may be required. A variety of methods have been described and new technologies are available. Cationic lipids, glycolipids, and fusion liposomes can also be used in the formulation (see, for example, WO 93/24640; Mannino & Gould-Fogerite, BioTechniques 6(7): 682 (1988); U.S. Patent No. 5,279,833 ; WO 91/06309; and Feigner et al, Proc. Nat'l Acad. Sci. USA 84:7413 (1987)). In addition, peptides and chemistries known as protective, interacting, non-agglomerating compounds (PINC) can also be complexed with purified plastid DNA to affect variables such as stabilization, intramuscular dispersion, or to specific organs or cells. The type of delivery. The stem cell sensitization can be used as a performance assay for the C TL epitope encoded by the minigene and a functional assay for the presentation of class I HLA. For example, plastid DNA is introduced into a mammalian cell line 142769.doc-120-201021828 suitable for use as a target for standard CTL chromium release assays. The method of transfection used will depend on the final formulation. Electroporation can be used for "naked" DNA, while cationic lipids allow direct transfection in vitro. The plastids expressing green fluorescent protein (GFP) can be co-transfected to allow for the enrichment of transfected cells using fluorescence activated cell sorting (FACS). The cells were then labeled with Col.-51 (51Cr) and used as a stem cell for the epitope-specific CTL line. 'The cytolysis based on the 5 Yr release rate indicates the production of the CTL epitope encoded by the minigene. Presented with hlA. The performance of HTL epitopes can be assessed in a similar manner using assays that assess HTL activity. In vivo immunogenicity is the second method of functional testing of small gene DNA formulations. A transgenic mouse expressing an appropriate human HLA protein is immunized with a DNA product. The dosage and route depend on the formulation (for example, intramuscular (iM) for DNA in PBS; intraperitoneal (for DNA complexed with lipids). The spleen cells are harvested on the first day after immunization, and The stimulation was further stimulated for one week in the presence of a peptide encoding each of the epitopes tested. After the treatment, CTL effector cells were assayed using standard techniques to assess the lysis of the loaded peptide, the standard cell of the 〇1* standard. HLA-sensitized target cell lysis with a peptide antigen (in response to a small gene-encoded epitope) demonstrates the DNA vaccine function of inducing CTL in vivo. The HTL epitope is demonstrated in a similar manner in the transgenic gene. Immunogenicity in mice. Alternatively, nucleic acids can be administered using a ballistic delivery method as described, for example, in U.S. Patent No. 5,204,253. The use of this technique to administer particles comprising only DNA. Adhere DNA to particles (such as gold particles) 142769.doc -121- 201021828. Small genes can also be transmitted using other bacteria or viral delivery systems well known in the art, for example, The expression construct of the epitope of the present invention is incorporated into a viral vector such as vaccinia. XC2. Combination of CTL peptide and accessory ribs 3. The plaque composition of the CTL peptide of the present invention can be modified (for example, by simulation) To provide the desired properties, such as a modified serum half-life, expanded population coverage, or enhanced leanness. For example, the peptide can be induced with at least one antigen capable of inducing a tau helper cell response. Determining the ligated linkage to enhance the ability of the peptide to induce CT1 activity. Although the CTL peptide can be directly ligated to the τ-assisted version, the CTL epitope/HTL epitope conjugate is typically ligated by a spacer molecule. This spacer is typically included in the physiology. a relatively small neutral molecule that is substantially uncharged under conditions, such as an amino acid or an amino acid mimetic. The spacer is typically selected, for example, from A1 a, Gly or a non-polar amino acid or a neutral polar amino acid. Other neutral spacers. It will be appreciated that spacers that are present as desired do not necessarily contain the same residues and may therefore be heterooligomers or homooligomers. When present, the spacers are typically at least 1 or 2 residues. , more usually 3 to 6 residues and sometimes 1 〇 or more than 1 residue. CTL peptide epitopes can be at (^[amino or terminal of the peptide, directly or via a spacer and T Auxiliary peptide epitope linkage. The immunogenic trait or T-helper amino terminus can be brewed. The HTL peptide epitope can also be modified to alter its biological properties. For example, it can be modified to include D- Amino acids enhance their resistance to proteases and thereby prolong their serum half-life, or they can bind to other molecules such as lipids 142769.doc-122-201021828, proteins, carbohydrates and their analogues to enhance their biology Activity. For example, a tau helper peptide can be linked to one or more palmitic acids at the amine or carboxy terminus. Combination of X.C.3 CTL Peptide and T Cell Sensitizer In some embodiments, it may be desirable to include at least one component that sensitizes B lymphocytes or T lymphocytes in the pharmaceutical compositions of the present invention. Lipids have been identified as agents capable of sensitizing CTL in vivo. For example, a cap acid residue can be attached to an epsilon-amino group and an a-amine group of an amine acid residue, and then, for example, via one or more linking residues (such as Gly, Gly-Gly_, Sei: , Sei^Sei· or a similar residue thereof) is linked to an immunogenic peptide. The lipidated peptide can then be administered directly as a microcell or granule incorporated into the liposome, or as an emulsion in an adjuvant (e.g., incomplete Freund's adjuvant). In a preferred embodiment, a particularly effective immunogenic composition comprises an ε-amino group attached to the ε-amino group of Lys and an α-amino group, which is linked to the immunogenic peptide via a linkage (Ser-Ser). The amine end. As another example of lipid sensitization of CTL reactions, E. coli lipoproteins such as tripalmitoyl-S-glyceryl cysteamine decyl syllidyl-serine (PsCSS) can be used in virus-specific CTL This CTL is sensitized when covalently linked to the appropriate peptide (see, eg, Deres et al, Nature 342:561, 1989). For example, a peptide of the present invention can be conjugated to P3CSS, and a lipopeptide can be administered to an individual to specifically sensitize an immune response against a target antigen. Furthermore, since the induction of neutralizing antibodies can also be sensitized with P3CSS-binding epitopes, the two compositions can be combined to more efficiently induce a humoral-mediated response and a cell-mediated response. 142769.doc -123- 201021828 XC4. Vaccine composition comprising DCs pulse-labeled with CTL and/or HTL peptides One example of a vaccine composition of the invention comprises administering a mixture of peptides having an epitope to the body in vitro PBMC from the patient's blood or DC isolated from it. A drug that is advantageous for harvesting DC can be used, such as ProgenipoietinTM (Pharmacia-Monsanto, St. Louis, MO) or GM-CSF/IL-4. DCs were washed to remove unbound peptides after pulsed DCs with peptides and before reinfusion into patients. In this embodiment, the vaccine comprises a peptide pulsed DC which presents a pulsed peptide epitope on its surface that is complexed with HLA molecules. DCs can be pulse-labeled ex vivo via peptide mixtures, some of which stimulate CTL responses to 58P1D12. A helper T cell (HTL) peptide that contributes to the CTL response, such as a natural or artificial loose restriction class II HLA peptide, may be included as needed. Thus, the vaccine of the present invention is useful for treating cancers that exhibit or overexpress 58P1D12. X.D.) Adoptive immunotherapy The antigenic 58P1D12-related peptide is also used to induce CTL and/or HTL responses ex vivo. The resulting CTL or HTL cells can be used to treat tumors in patients who do not respond to other conventional therapeutic modalities or who do not respond to the therapeutic vaccine peptides or nucleic acids of the invention. Induction of specific antigens by culturing a patient or a genetically compatible CTL or HTL precursor cell together with an antigen presenting cell (APC) (such as a dendritic cell) source and a suitable immunogenic peptide in tissue culture CTL or HTL reaction. After cultivating for an appropriate period of time (usually about 7-28 days) (in which the precursor cells are activated and propagated into effector cells), the cells are returned to the affected area 142769.doc -124-201021828, thereby destroying their specific markers. Cells (eg, tumor cells) (CTLs) or promote the destruction of their specific target cells (eg, tumor cells) (HTL). Transfected dendritic cells can also be used as antigen presenting cells. X · E.) Administration of Vaccines for therapeutic or prophylactic purposes The pharmaceutical and vaccine compositions of the present invention are generally used to treat and/or prevent cancers that exhibit or overexpress 5 8P1D12. In therapeutic applications, the peptide and/or nucleic acid composition is administered to a patient in an amount sufficient to elicit an effective B cell, CTL and/or HTL response against the antigen and sufficient to cure or at least partially arrest or reduce symptoms and/or complication. An amount sufficient to accomplish this is defined as a "therapeutically effective dose." The amount effective for this use will depend, for example, on the particular composition being administered, the mode of administration, the stage and severity of the condition being treated, the weight and general condition of the patient, and the judgment of the attending physician. For pharmaceutical compositions, the immunogenic peptides of the invention or DN A encoding the same are typically administered to an individual who already has a tumor that exhibits 5 8P1D12. The DNA in which the ORF or φ is encoded may be administered alone or in the form of a fusion of one or more peptide sequences. The patient may be treated with an immunogenic peptide, alone or in combination with other treatments, such as surgery, as appropriate. For therapeutic use, administration should usually begin on the first diagnosis of a 58 piD 相 2 related cancer. The dose is then increased until the symptoms are at least substantially relieved and maintained for a period of time. Examples of vaccine compositions delivered to a patient (i.e., including, but not limited to, examples of peptide mixtures, multi-antigenic polypeptides, minigenes or TAA-specific CTLs or pulse-labeled dendritic cells) Change according to the stage of the disease or the health status of the patient. For example, 142769.doc 125-201021828 A vaccine comprising a 58P1D12-specific CTL in a patient with a tumor exhibiting 58P1D12 can more effectively kill tumor cells in patients with advanced disease compared to alternative embodiments. . It is generally important to provide an amount of a peptide epitope that is delivered by a mode of administration sufficient to effectively stimulate a cytotoxic T cell response; a composition that stimulates a helper T cell response can also be obtained in accordance with this embodiment of the invention. The dose of initial therapeutic immunization typically occurs in a unit dosage range with a lower t limit of about i, 5, 50, 500 or 1000 and an upper limit of about 10,000, 2 〇' 〇〇〇, 30,000 or 50,000 ton. Human dose values typically range from about 500 pg to about 50,000 calls per 70 kg patient. As determined by measuring the specific activity of CTL and HTL obtained from the patient's blood, depending on the patient's response and condition, it can be administered from about 1.0 gg to about 50 in weeks to months according to intensive therapy. Call between the peptide fortification dose. Administration should continue until the clinical symptoms or laboratory tests indicate that neoplasia has been eliminated or reduced and this condition is maintained for a period of time. The dosage, route of administration, and duration of administration are adjusted according to methods known in the art. In certain embodiments, the peptides and compositions of the invention are used in a critical condition «i.e., a life-threatening or potentially life-threatening condition. In such cases 'because the amount of foreign material is extremely small and the peptides of the preferred compositions of the invention are relatively non-toxic', it is possible to administer the peptide compositions in such amounts as the substantial amount of such peptides administered and treated The physician may feel that this is necessary. The vaccine composition of the present invention may also be used only as a prophylactic dose. The dose of the immunization usually occurs in a unit dose range, and the #lower limit value is about 1, 5, 50, 500 or 1000 μ§ and the upper limit is about 1 〇, _, 2〇, _, 142769.doc -126- 201021828 30,000 or 50,000 pg. Human dose values typically range from about 500 pg to about 50,000 pg per 7 kg of patient. Thereafter, about 4 to 6 months after the initial administration of the vaccine, a fortified dose of peptide between about 1 〇 to about 5 〇, 〇〇〇 投 is administered at defined intervals. The immunogenicity of the vaccine can be assessed by measuring the specific activity of CTL and HTL obtained from the patient's blood sample. Pharmaceutical compositions for therapeutic treatment are intended to be administered parenterally, topically, orally, nasally, intrathecally or topically (e.g., in the form of a cream or topical ointment). The pharmaceutical composition is preferably administered parenterally (e.g., intravenous, subcutaneous, intradermal or intramuscular). Accordingly, the present invention provides a composition for parenteral administration comprising a solution in which an immunogenic peptide is dissolved or suspended in an acceptable carrier, preferably an aqueous carrier. A variety of aqueous carriers can be used, such as water, buffered water '〇 8% saline, 0.3% glycine, uronic acid, and the like. The compositions may be sterilized by conventional, well known sterilization techniques or may be sterile filtered. The resulting aqueous solution may be packaged for use as is or lyophilized, and the lyophilized preparation is combined with a sterile solution prior to administration.组合 The composition may contain pharmaceutically acceptable auxiliary substances required for physiological conditions, such as pH adjusting agents and buffers, tonicity adjusting agents, sizing agents, preservatives and the like, such as sodium acetate, sodium lactate, Gasified steel, potassium carbonate, calcium chloride, sorbitan monolaurate, oleic acid triester, and the like. The concentrations of the peptides of the present invention in pharmaceutical formulations can vary widely, i.e., are small; about 1 weight/twist, typically about 2% by weight or at least about 2% by weight up to 20% to 50% by weight. Set /. The second time is more than 5 ,, and mainly based on the fluid body 142769.doc ·] 27· 201021828 Product, viscosity, etc. are selected according to the specific drug delivery mode selected. Pharmaceutical compositions typically comprise a composition in a human unit dosage form comprising a human unit dose of an acceptable carrier (in one embodiment, an aqueous carrier) and known to those skilled in the art to The composition is again ', human volume / volume contribution (see for example (10), s harmaceutical Sciences '17th edition, edited by A. Gennaro,] VJack

Publishing Co., Easton, Pennsylvania, 1985). For example, the peptide dose for initial immunization may range from about 1 to about 50,000 for a 7 〇 kg patient' typically 1 〇〇 5 〇〇〇 (d). For example, for nucleic acids, the initial immunization can be performed in an amount of _5_5 mg using a performance vector in a naked nucleic acid administered intramuscularly (or subcutaneously or intradermally) at a plurality of sites. It is also possible to use a gene-scraping and nucleic acid (〇1 to 1000 pg) ^ 3_4 weeks incubation period followed by a booster dose. The enhancer may be a recombinant fowlpox virus administered at a dose of 5 < 1 〇 7 to 5 x 1 〇 9 pfu. Treatment with antibodies typically involves repeated administration of an anti-58P1D12 antibody formulation via an acceptable route of administration (such as intravenous injection (IV)), typically at a dose ranging from about 0_1 to about 10 mg/kg body weight. In general, doses in the range of 10-500 mg MAb per week are effective and well tolerated. In addition, an intravenous initial loading of about 4 mg/kg of patient body weight of the anti-58P1D12 MAb formulation, followed by an intravenous dose of about 2 mg/kg per week, represents an acceptable dosage regimen. As will be appreciated by those skilled in the art, a variety of factors can affect the desired dosage in a particular situation. Such factors include, for example, the half-life of the composition, the binding affinity of the Ab, the immunogenicity of the substance, the degree of 58P1D12 expression in the patient, the extent of circulating the 58P1D12 antigen, the desired 142769.doc •128-201021828 steady state concentration, the frequency of treatment And the effects of a chemotherapeutic or other agent in combination with the method of treatment of the invention, as well as the health status of a particular patient. Non-limiting preferred human unit doses are, for example, 5 〇〇pg-i mg, 1 mg-50 mg, 50 mg-100 mg, 100 mg-200 mg, 200 mg-300 mg, 400 mg-500 mg, 500 mg -600 mg, 600 mg-700 mg, 700 mg-800 mg, 800 mg-900 mg, 900 mg-1 g or 1 mg-700 mg. In certain embodiments, the dosage is, for example, in the range of 2-5 mg per kilogram of body weight, followed by a weekly dose of 1-3 mg/kg; for example, 5 mg, 1 mg, 2 mg, 3 mg per kg body weight , 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg followed by a weekly dose of two weeks, three weeks or four weeks; for example 0.5-10 mg per kg body weight, followed by two Weekly, three or four weeks weekly dose; body area of 225, 250, 275, 300, 325, 350, 375, 400 mg m2 per week; body area of 1-600 mg m2 per week; 225-400 mg per week The body area of m2; the dose may be followed by a weekly dose of 2, 3, 4, 5, 6, 7, 8, 9, 19, 11, 12 or more weeks. In one embodiment, the human unit dosage form of the polynucleotide comprises an appropriate dosage range or effective amount to provide any therapeutic effect. As will be appreciated by those of ordinary skill, the therapeutic effect will depend on a number of factors, including the sequence of the polynucleotide, the molecular weight of the polynucleotide, and the route of administration. The dosage is typically selected by a physician or other health care professional based on a variety of parameters known in the art, such as severity of the condition, patient history, and the like. In general, for polynucleotides having about 20 bases, the dosage range can be selected, for example, from, for example, about 0.1, 0-25, 0.5, 1, 2, 5, 10, 20, 30, 40, 50, 60, 70. , 80, 90, 100, 200, 300, 400 or 500 mg/kg 142769.doc -129- 201021828 The lower limit of independent selection up to approximately 60, 80, 100, 200, 300, 400, 500, 750, 1000 ' The upper limit of the independent selection of 1500, 2000, 3000, 4000, 5000, 6000, 7000 '8000, 9000 or 10,000 mg/kg is greater than this lower limit. For example, the dose can be about any of the following: 0.1 to 100 mg/kg, 0.1 to 50 mg/kg, 0.1 to 25 mg/kg, 0.1 to 10 mg/kg, 1 to 500 mg/kg, 100 to 400 Mg/kg, 200 to 300 mg/kg, 1 to 100 mg/kg, 100 to 200 mg/kg, 300 to 400 mg/kg, 400 to 500 mg/kg, 500 to 1 mg/kg, 500 To 5000 mg/kg or 500 to 1 〇, 〇〇〇mg/kg. In general, a parenteral route of administration may require a higher dose of a polynucleotide, such as an increased length of polynucleotide, as compared to applying the nucleotide directly to the diseased tissue. In one embodiment, the human unit dosage form of the T cell comprises an appropriate dosage range or effective amount to provide any therapeutic effect. As the average person understands, the therapeutic effect depends on a number of factors. The dosage is typically selected by a physician or other health care professional based on a variety of parameters known in the art, such as the severity of the condition, the patient's medical history, and the like. The dose may be from about 1 to 4 cells to about 1 to 6 cells, from about 1 to 6 cells to about 1 to 8 cells, from about 1 to 8 to about 6 to about 11 cells or from about 1 to about 8 to about 5 Hey, 0 cells. The dose may also range from about 1 to 6 cells per square meter to about 1 to 10 cells per square meter, or from about 106 cells per square meter to about 1 to 8 cells per square meter. The protein of the present invention and/or the nucleic acid encoding the protein may also be administered via a liposome, and the liposome may also be used to: select the protein core to the special 'heart lymphatic tissue; 2) selectively (4) to the diseased cell; or 3) Extend the half-life of the peptide composition. Liposomes include emulsions, foams, micelles, 142769.doc • 130- 201021828 soluble monolayers, liquid crystals, phospholipid dispersions, flaky layers and the like. In such preparations, the peptide to be delivered alone or together with a molecule that binds to a receptor prevalent in lymphoid cells, such as a monoclonal antibody that binds to a CD45 antigen, or together with other therapeutic or immunogenic compositions Combined as part of the liposome. Thus, liposomes filled or modified by the desired peptide of the present invention can be directed to the site where the lymphoid cells are located, wherein the liposomes then deliver the peptide composition. Lipid systems for use in accordance with the present invention are formed from standard liposome-forming lipids. These lipids typically include neutral and negatively charged fats and sterols, such as cholesterol. Lipid selection is typically guided by consideration of, for example, liposome size, acid instability and stability of the liposome in the bloodstream. A variety of methods for preparing liposomes can be used, as described in, for example, Sz〇ka et al, Ann. Rev. Biophys. Bioeng. 9:467 (1980); and U.S. Patent Nos. 4,235,871, 4,501,728 , 4, 837, 028 and 5, 〇 19, 369. For the donor cell of the immune system, the ligand to be incorporated into the liposome may include, for example, an antibody or a fragment thereof specific for the cell surface determinant of the desired immune system cell. The liposome suspension containing the peptide can be administered intravenously, locally, topically, etc., depending on the mode of administration, the peptide delivered, and the stage of the disease being treated. For solid compositions, conventional non-toxic solid carriers can be employed, including, for example, pharmaceutical grades of mannitol 'lactose, starch, stearic acid, sodium saccharin, talc, cellulose, glucose, Yan. Sugar, carbonated towns and their analogues. For oral administration, any of the usual excipients (such as those listed above) can be used with the usual! 〇 95% (and more preferably 25% _75% 142769.doc • 131· 201021828 peptide) combined active ingredient (ie, one or more pharmaceutically acceptable non-toxic combinations of the invention) For aerosol administration, the immunogenic peptide is preferably provided in the form of a fine powder along with the surfactant and the propellant. A typical percentage of the peptide is about 8% by weight, preferably about one weight (four). -10% by weight. Of course, the surfactant must be non-toxic and preferably soluble in the propellant. These agents are represented by fatty acids containing from about 6 to 22 carbon atoms (such as caproic acid, octanoic acid, lauric acid, Brown acid, stearin, linoleic acid, linoleic acid, olesteric acid and oleic acid) and aliphatic polyols or vinegars or partial esters thereof. Mixed esters such as a glyceride or a natural glyceride. The surfactant may comprise from about 5% by weight to about 2% by weight of the composition, preferably from about 0.25 to 5% by weight. The remainder of the composition is typically a propellant. For example, for the intranasal delivery of lecithin. XI) 58P1D1 Diagnosis and Prognosis of 2 As disclosed herein, 58P1D12 polynucleotides, polypeptides, reactive cytotoxic T cells (CTL), reactive helper tau cells (HTL), and anti-polypeptide antibodies can be used to examine changes associated with cell growth disorders. Well-known diagnostic, prognostic, and therapeutic assays for conditions such as cancer, particularly those listed in Table I (see, for example, specific patterns of tissue performance and overexpression in certain cancers, such as for example the title " 58P1D12 is described in the example of performance analysis in normal tissues and patient samples). 58P1D12 can be similar to the ovarian cancer-associated antigen CA125, a biomarker used by medical practitioners to identify and monitor the presence of ovarian cancer for many years (see, eg, Gagnon and Ye, Curr. Opin· Obstet. Gynecol. 2008; 142769.doc • 132· 201021828 20:9-13). A variety of other diagnostic markers can be used in similar contexts, including p53 and K-ras (see, for example, Tulchinsky et al., int j Mol Med July 4 (1): 99-102 and Minimoto et al., Cancer Detect Prev 2000 ; 24(1): 1-12). Thus, the disclosure of 58P1D12 polynucleotides and polypeptides (and 58P1D12 polynucleotide probes and anti-58P1D12 antibodies for identifying the presence of such molecules) and their properties allows those skilled in the art to These molecules are used in a similar manner to the methods used in examining various diagnostic assays for cancer-related conditions. A typical example of a diagnostic method using 5 8P1D12 polynucleotides, polypeptides, reactive tau cells, and antibodies is similar to the use of well established diagnostic assays such as PSA polynucleotides, polypeptides, reactive T cells, and antibodies. These methods. For example, as PSA polynucleotides can be used as probes (for example in Northern analysis, see, for example, Sharief et al, Biochem. Mol. Biol. Int. 33(3): 567-74 (1994)) and Primers (for example, in PCR assays, see, for example, Okegawa et al, J. Urol. 163(4): 1 189-1190 (2000)) to observe PSA mRNA in a method for monitoring PSA overexpression or metastasis in prostate cancer. The presence and/or amount of the 58P1D12 polynucleotide described herein can be used in the same manner to detect 58P1D12 overexpression or metastasis in ovarian cancer and other cancers that exhibit this gene. Alternatively, just as PSA polypeptides can be used to generate antibodies specific for PS A, such antibodies can then be monitored for excessive expression of PSA proteins (see, eg, Stephan et al, Urology 55(4):560-3 (2000)) or prostate cells. For the transfer (see, for example, Alanen et al, Pathol. Res. Pract. 192(3): 233-7 (1996)) for the observation of the presence and/or amount of PSA protein, 142769.doc-133 described herein. - 201021828 The 5 8P 1D12 polypeptide can be used to produce anti-specific expression or metastasis of 58P1D12 in cells for detecting ovarian cells and other cancers exhibiting this gene, since metastasis includes cancer cells from the source organ (such as the lungs) Or ovaries #) move to different areas of the body (such as lymph nodes), so evidence of transfer can be provided using a test to check for the presence of cells expressing 58P1D12 polynucleotides and/or polypeptides in a biological sample. For example, when a biological sample that normally does not contain tissue of 58P1D12 expressing cells contains 58 piD12 expressing cells, this finding indicates metastasis. Or '58P1D12 poly-picuric acid and/or polypeptide can be used to provide evidence of cancer, for example, when a cell in a biological sample that does not exhibit 58P1Di2 under normal conditions or exhibits 58P1D12 to varying degrees exhibits 58P1D12 or has a stronger expression of 58P1D12 (See, for example, the 58P1D12 performance in the cancer samples listed in Table I and in the patient samples shown in the figures). In such assays, those skilled in the art may further desire to generate additional evidence of the transfer by testing whether the biological sample A has a second tissue restriction marker (other than 5 8 P1D12). The use of immunohistochemistry to identify the presence or absence of a 58P1D12 polypeptide in a tissue section can indicate a change in the state of certain cells within the tissue. It is well understood in the art that the ability of an antibody to localize to a polypeptide expressed in a cancer cell is a diagnostic disease, stage of disease, development, and/or tumor invasiveness. This antibody also detects changes in the distribution of polypeptides in cancer cells compared to corresponding non-malignant tumor tissues. 5 8P1D12 polypeptides and immunogenic compositions are also suitable for observing disease states. 142769.doc -134- 201021828 The phenomenon of subcellular protein localization changes. Changes in cells from normal to disease state result in changes in cell morphology and are usually accompanied by changes in subcellular protein localization/distribution. For example, cell membrane proteins that are expressed in a polarized manner in normal cells can change in disease, resulting in a protein that is distributed non-polarly throughout the cell surface. The phenomenon of subcellular protein localization changes under disease conditions has been demonstrated by the use of immunohistochemical methods via MUC 1 and Her2 protein expression. In addition to some nuclear localization of glycoproteins, normal epithelial cells also have a typical apical distribution of MUC 1 ® , while malignant lesions often exhibit a non-polar staining pattern (Diaz et al., The Breast Journal, 7; 40-45 (2001) Zhang et al, Clinical Cancer Research, 4; 2669-2676 (1998); Cao et al, The Journal of Histochemistry and Cytochemistry, 45: 1547-1557 (1997)). In addition, normal breast epithelium is negative for Her2 protein, or only shows basolateral distribution, while malignant cells can express this protein on the entire cell surface (De Potter et al, International Journal of Cancer, 44; 969-974 (1989) McCormick et al., 117; 935-943 (2002)). Alternatively, the distribution of the protein can vary from surface location only to include diffuse cytoplasmic manifestations under diseased conditions. This example can be found in MUCl (Diaz et al, The Breast Journal, 7: 40-45 (2001)). Changes in the location/distribution of proteins in cells, as detected by immunohistochemistry, can also provide important information about the convenience of certain therapeutic modalities. Finally, this point is illustrated by the fact that the protein can be located in the cells in normal tissues but on the cell surface in malignant cells. 142769.doc -135- 201021828 Cell surface localization makes the cells beneficial for antibody-based diagnosis and treatment. Treatment. The 58P1D12 protein and its associated immune response are extremely useful when this change in protein localization occurs for 58P1D12. The use of the 5 8P1D12 composition allows those skilled in the art to make important diagnostic and therapeutic decisions. When a polypeptide is present in a tissue that does not normally produce 58P1D12, the immunohistochemical reagent specific for 58P1D12 can also be used to detect metastasis of a tumor exhibiting 58P1D12. Thus, the 58P1D12 polypeptide and antibodies produced by its immune response can be used in a variety of important settings, such as diagnostic, prognostic, prophylactic, and/or therapeutic purposes known to those skilled in the art. Furthermore, the 58P1D12-related protein or polynucleotide of the present invention can be used to treat a pathological condition characterized by overexpression of 58P1D12. For example, the amino acid sequence or nucleic acid sequence of Figure 1 or a fragment of either can be used. To generate an immune response against the 58P1D12 antigen. An antibody or other molecule that reacts with 58P1D12 can be used to modulate the function of this molecule and thereby provide a therapeutic benefit. XI.A.) Inhibition of 58P1D12 Protein Function The present invention includes various methods and compositions for inhibiting the binding of 58P1D12 to its binding partner or inhibiting the binding of 58P1D12 to other proteins, as well as methods for inhibiting the function of 58P1D12. XI.B.) Inhibition of 58P1D12 via intracellular antibodies In one method, a recombinant vector encoding a single-chain antibody that specifically binds to 58P1D12 is introduced into 58P1D12 expressing cells via gene transfer technology 142769.doc • 136·201021828. Thus, the encoded single-stranded anti-58P1D12 antibody is expressed in the cell to bind to the 5 8 P1D12 protein and thereby inhibit its function. Methods for engineering such single-chain antibodies in cells are well known. These intracellular antibodies (also referred to as "intracellular antibodies") are specifically directed to specific metabolic regions within the cell to provide control at the concentration of inhibitory therapeutic activity. This technique has been successfully applied in this technology (for a review of 's Qing, see Richardson and Marasco, 1995, TIBTEC, Vol. 13). Intracellular antibodies have been shown to substantially eliminate the expression of other abundant cell surface receptors (see, for example, Richards〇n et al., ® I" 5, Pro. Natl. Acad. Sci. USA 92: 3137-3 141; Beerli et al. Human, 1994, J. Biol. Chem. 289: 23931-23936; Deshane et al. '1994, Gene Ther. 1: 332-337). Single-chain antibodies comprise the variable domains of the heavy and light chains joined by a flexible linker polypeptide and are expressed as a single polypeptide. The single bond antibody is expressed as needed in the form of a single-chain variable region fragment joined to the light chain constant region. Well-known intracellular signaling signals are engineered into recombinant parameric nucleotide vectors encoding such single chain antibodies to allow intracellular antibodies to be accurately targeted to the desired intracellular metabolic region. For example, intracellular antibodies targeting the endoplasmic reticulum (ER) have been engineered

The wKDEL amino acid motif is combined with a leader peptide and optionally a C-terminal ER indwelling signal. Intracellular antibodies designed to be active in the nucleus are engineered to include nuclear localization signals. The lipid moiety is conjugated to the intrabody to retain the intracellular resistance system to the cytosol side of the plasma membrane. Intracellular antibodies can also be targeted to function in cytosol. For example, cytosolic intracellular antibodies are used to sequester a factor into a cytosol to prevent its transport to its natural cell end point. 142769.doc 137- 201021828 In a consistent application, intracellular antibodies were used to capture 58P1D12 in the nucleus, thereby preventing its activity in the nucleus. Nuclear targeting signals are engineered into these 5<8>P1D12 intracellular antibodies' to achieve the desired orientation. These 58ρΐΓ)12 intrabodies are designed to specifically bind to a specific 58Ρ1Γ12 domain. In another embodiment, a cytosolic intracellular antibody that specifically binds to the 58P1D12 protein is used to prevent 58P1D12 from reaching the nucleus, thereby preventing it from exerting any biological activity in the nucleus (e.g., preventing 58P1D12 from forming a transcription complex with other factors). XI.C.) Inhibition of 58P1D12 via Recombinant Protein In another method, the recombinant molecule binds to 58P1Di2 and thereby inhibits 58 P1D12 function. For example, the recombinant molecules prevent or inhibit 58P1D12 from approaching/binding its binding partner or binding to other proteins. Such recombinant molecules may, for example, contain a reactive portion of a 58P1D12 specific antibody molecule. In a specific embodiment, the 58P1D12 binding partner's 58P1D12 binding domain is engineered into a dimeric fusion protein, wherein the fusion protein comprises two 58P1D12 ligands linked to the Fc portion of a human IgG (such as human IgG丨) Body binding domain. This IgG portion may contain, for example, the CH2 and CH3 domains and the hinge region' without the C% domain. This dimeric fusion protein is administered in a soluble form to a patient suffering from a cancer associated with 58P1D12, wherein the dimeric fusion protein specifically binds to 58P1D12 and blocks the interaction of 58 P1D12 with the binding partner. The dimeric fusion proteins are further combined into a multimeric protein using known antibody ligation techniques. XI.D.) Inhibition of 58P1D12 Transcription or Translation 142769.doc .138· 201021828 The present invention also encompasses various methods and compositions for inhibiting the transcription of the 58P1D12 gene. Similarly, the invention also provides methods and compositions for inhibiting the translation of mRNA into a protein. In one method, a method of inhibiting transcription of the 58P1D12 gene comprises contacting a 58P1D12 gene with a 58P1D12 antisense polynucleotide. In another method, a method of inhibiting translation of 58P1D12 mRNA comprises contacting 58P1D12 mRNA with an antisense polynucleotide. In another method, the 58P1D12 specific ribozyme split 581 > 1012 message is used to inhibit translation. Such antisense and ribozyme based methods can also be directed to the delta-circumscribed region of the 58P1D12 gene such as the 58P1D12 promoter and/or enhancer element. Similarly, a protein capable of inhibiting the 58P1D12 gene transcription factor was used to inhibit the transcription of 58P1D12 mRNA. A variety of polynucleotides and compositions suitable for use in the above methods have been described above. The use of antisense and ribozyme molecules to inhibit transcription and translation is well known in the art. Other factors that inhibit the transcription of 58P1D12 by interfering with 58P1D12 transcriptional activation are also suitable for the treatment of cancers that exhibit 58P1D12. Similarly, the factor that interferes with 58P1D12 processing is suitable for treating cancers that exhibit 58 piD12. Cancer treatment methods using these factors are also within the scope of the invention. ΧΙ·Ε.) For the general consideration of therapeutic strategies, gene transfer and gene therapy techniques can be used to treat therapeutic polynucleotide molecules (ie, antisense and ribozyme polynucleotides encoding intracellular antibodies and other 58P1D12 inhibitory molecules). Transfer to tumor cells that synthesize 58P1D12. A variety of gene therapy methods are known in the art. Using these gene therapy methods, a heavy material encoding a 58P1D12 antisense polynucleotide, a nuclear 142769.doc-139-201021828 enzyme, and a factor capable of interfering with 58P1D12 transcription can be delivered to the target m cells. Any combination of surgical chemotherapy or radiation therapy. The present m treatment allows for low dose chemotherapy (or other therapy) and/or fewer doses, which is not good for all patients and especially for Patients who are resistant to chemotherapeutic toxicity are _ advantages. The anti-tumor activity of the sputum group (such as antisense molecules, ribozymes, intracellular antibodies) or a combination of these sites can be used in a variety of in vitro and in vivo Verification of the assay. In vitro assays for assessing therapeutic activity include cell growth assays, soft agar assays, and other assays for the expression of the tumor-promoting activity. A binding assay that determines the extent to which a therapeutic composition inhibits the binding of 58P1D12 to a binding partner. Et al. The in vivo effects of the 58P1D12 therapeutic composition can be assessed using an appropriate animal model. For example, a heterogeneous ovarian cancer model can be used, in which a person That is, nested cancer explants or secondary xenograft tissues are introduced into immunocompromised animals, such as nude mice or SCID mice (Klein et al, 1997, Nature Medicine 3: 402-408). For example, PCT patent application W〇〇98/16628 and U.S. Patent 6,107,540 describe various xenograft models of human prostate cancer that can reproduce the appearance of primary tumors, micrometastasis, and the formation of osteoblastic features of advanced disease. A test for predicting tumor formation inhibition, tumor regression or metastasis, and the like, predicts efficacy. An in vivo assay to assess accelerated apoptosis is suitable for assessing therapeutic compositions. In one embodiment, therapeutic properties can be examined. Whether the xenograft A of the mice treated with the composition treated with the 142769.doc-140-201021828 tumor had a focal focus of apoptosis and compared with untreated control mice with xenografts. The extent to which the focus of apoptosis is present in the tumor indicates the therapeutic efficacy of the composition. The therapeutic composition used to practice the above methods can be formulated to include A pharmaceutical composition for delivering a carrier of a method. Suitable carriers include any material that, when combined with a therapeutic composition, retains the antitumor function of the therapeutic composition and is generally non-reactive with the patient's immune system. Examples include (but not Limited to any of a variety of standard pharmaceutical carriers, such as sterile saline buffer aqueous buffer solutions, bacteriostatic water and the like (see generally Remingt〇n, s Pharmaceutical Sciencea 16 edition, Α·〇sal. ed., i98 The therapeutic formulation is soluble and can be administered via any route that is capable of delivering the therapeutic composition to the tumor site. Potentially effective routes of administration include, but are not limited to, intravenous, parenteral, intraperitoneal, intramuscular, intratumoral, intradermal, intraorganic, orthotopic, and the like. Preferred formulations for intravenous injection include sterile water dissolved in antiseptic treatment, sterile water without preservative treatment, and/or diluted in a mixture containing 〇.9% sterile sodium chloride for injection (usp) A therapeutic composition in a vinyl chloride or polyethylene bag. The therapeutic protein preparation may be lyophilized and preferably stored under vacuum in the form of a sterile powder and then reconstituted in bacteriostatic water (containing, for example, a benzyl alcohol preservative) or reconstituted in sterile water. Dosages and dosing regimens for treating cancer using the above methods vary with the method and target cancer, and are generally dependent on a variety of other factors known in the art. ', XII.) Identification, Characterization and Use of 58P1D12 Modulators 142769.doc • 141 - 201021828 Method of Monitoring and Using Regulators In one example, 'screening is performed to identify induction or inhibition of specific expression distribution, inhibition or induction of specificity A modulator of a sexual route, preferably resulting in a related phenotype. In another embodiment, screening is performed to identify modulators that alter (increase or decrease) the performance of individual genes by identifying differentially important genes that are important in a particular state. In another embodiment, screening is performed to identify modulators that alter the biological function of the expression product of the differentially expressed gene. Furthermore, by identifying genes that are important in a particular state, screening is performed to identify agents that are biologically active, 纟α-and/or modulate the biological activity of the gene product. In addition, screening is performed on genes induced in response to the candidate agent. Screening is performed to identify genes that are specifically regulated in response to the agent, after identifying modulators (inhibiting the pattern of cancer expression to produce a normal expression pattern of the modulator, or causing the gene to behave like a general cancer gene modulator in normal tissues). Performance distribution comparison between normal tissue and drug-treated cancer tissue 'Discloses genes that are not exhibited in normal tissues or cancer tissues but are expressed in tissues treated with the drug, and vice versa. Such agent-specific sequences are identified and used by the methods described herein for cancer genes or proteins. In particular, the sequences and the proteins encoded thereby are used to label or clock the cells treated with the agent. In addition, antibodies are produced against the agent-inducing proteins, and such antibodies are used to target novel therapeutic agents to a treated cancer tissue sample. Regulator-related screening and screening assays: Gene performance-related assays Proteins, nucleic acids, and antibodies of the invention are used in screening assays. Screening 142769.doc -142- 201021828 The use of cancer-associated proteins, antibodies, nucleic acids, modified proteins, and cells containing such sequences, such as assessing the "gene expression distribution", polypeptide distribution, or biological function of a candidate drug The impact of change. In one embodiment, a preferred combination high throughput screening technique uses a performance profile to allow monitoring of performance profile genes after treatment with a candidate agent (eg, DaWs, GF et al, j Biol Screen 7:69 (2〇) 〇2) ; 21〇1^仙 et al, Science 279:84_8 (1998) ; Heid, Gen_ ^ 6:986 94, 1996) 〇, screening proteins used in cancer proteins, antibodies, nucleic acids, modified proteins And cells containing natural or modified cancer proteins or genes. That is, the present invention encompasses a method of screening a composition for modulating the cancer phenotype or physiological function of the cancer protein of the present invention. This can be performed against the gene itself or by assessing the effect of the candidate drug on the "gene expression profile" or biological function. In one embodiment, it is preferred to use a performance profile in conjunction with a high pass (four) test technique to allow Is measurements to be taken after treatment with a candidate agent, see Zlokamik, supra. A variety of assays are performed for the genes and proteins of the invention. The assay is carried out at the individual white matter of the white protein, via a specific gene that is up-regulated in cancer, and the test compound is tested for its ability to modulate gene expression or to bind to the cancer protein of the present invention. "Adjustment" in this context includes an increase or decrease in the performance of the gene. The preferred amount of adjustment depends on the initial change in the expression of the gene in the normal tissue versus the tissue undergoing the cancer, with a change of at least 10/. Preferably, 50/〇, more preferably 10〇_3〇〇% 'and in some embodiments 3〇 (M_% or greater than 1000%. Therefore, if compared to normal tissue, base 142769.doc -143- 201021828 Because it shows a 4-fold increase in cancer tissue, it usually takes about four times the reduction; similarly, if the gene is reduced by 10 times in cancer tissue compared to normal tissue, then the performance of the test compound usually requires 1G doubling. Target value. Regulators that can be used to aggravate gene expression types in cancer can also be used, for example, as targets for up-regulation in further analysis. Use of nucleic acid probes and quantified gene expression to quantify the amount of performance of the county, or for example The gene product itself is monitored by using antibodies against cancer proteins and standard immunoassays. Proteomic research and isolation techniques also allow for quantification of performance. Monitoring of the performance of compounds that regulate gene expression in the examples is monitored in the example of ' |5J The monitoring of the gene expression of an entity, that is, the distribution of performance. These distributions usually include one or more of the bases of the graph. The needle is attached to the biochip to measure and quantify the cancer sequence in a particular cell. Alternatively, PCR can be used. Thus, the primer can be dispensed into a predetermined well using, for example, a microtiter plate. The PCR reaction can then be performed and the wells can be analyzed. Performance monitoring is performed to identify compounds that modulate the expression of one or more cancer-related sequences (eg, the poly(tetra) acid sequence shown in Figure:). Typically, the assay is added to the cells during knifeing. Also provided are screening methods for identifying cancer, modulating a cancer protein of the invention, binding to a cancer protein of the invention, or interfering with a pharmaceutical protein of the invention in combination with an antibody or other binding partner. In one embodiment, The high-throughput screening method includes providing a library containing a large number of potential therapeutic wow compounds (candidate compounds). The "combined chemical libraries" are then examined by one or more assay screens 142769.doc 201021828 to identify the ones that exhibit the desired characteristic activity. Such library members (specific chemical species or subclasses identified by this compound can be used as a conventional "dominant compound", screening compound Or a therapeutic agent. In certain embodiments, screening a combinatorial library of potential modulators is capable of binding to a cancer polypeptide or modulating activity. Conventionally, by identifying a compound having certain desired properties or activities (eg, inhibitory activity) (referred to as "dominant compounds"), produce variants of dominant compounds and evaluate the properties and activities of their variant compounds to produce novel chemicals with useful properties. This analysis typically uses high-throughput screening (HTS) Method As described above, gene expression monitoring can be conveniently used to test for candidate modulators (eg, protein f, nucleic acid or small molecule). Adding a candidate agent and incubating the cells for a period of time will, for example, contain the stem sequence to be analyzed The sample is added to the biochip. The target sequence is prepared using known techniques as needed. For example, the sample is treated with a known lysis buffer, electroporation, or the like to lyse the cells, where purification and/or amplification, such as pCR, is performed as appropriate. For example, in vitro transcription is performed by a label covalently linked to nucleotide acid.胄 Commonly used biotin-FITC or PE or labeled with Cy3 or cy5. The stem sequence can be labeled, for example, with a fluorescent signal, a chemiluminescent signal, a chemical signal, or a radioactive signal to provide a means by which the valence-labeled stem sequence specifically binds to the probe. The standard can also be an enzyme, such as an alkaline phosphatase or horseradish peroxidase, which, when provided with the appropriate quality, produces a detectable product. Alternatively, labeled compounds or small molecules, such as enzyme inhibitors, that are labeled but not catalyzed or altered by an enzyme. The label may also be a moiety or compound that specifically binds to the anti-protein chain 142769.doc • 145· 201021828, such as an epitope tag or biotin. Taking biotin as an example, streptavidin is labeled as described above to provide a detectable signal for the binding sequence without sequence. Unbound labeled anti-streptavidin is usually removed prior to analysis.

As will be appreciated by those skilled in the art, such assays may be direct hybridization assays or may include "sandwich assays" which include the use of multiple probes as generally outlined in the following U.S. patents: 5, 6817 2, 5'597, 909, 帛5,545,730, 帛5,594, ιΐ7, 5,591,584, 帛5,571,67〇, 帛5 58〇, 731, 5,57^70, 帛5,591 , No. 584, 帛 5, 624, Xie, No. 5, 635, 352, 帛 5, 594, 118, 帛 5 359 1 第 No. 5, 124, 246 and No. 5, 681, 697. In this implementation, in general, silk nucleic acids are prepared as described above and then added to a biochip comprising a plurality of nucleic acid probes under conditions permitting formation of a hybridization complex. The invention employs a variety of hybridization conditions, including highly moderate and low stringency conditions as described above. The verification material is carried out under strict conditions, such

Stringent conditions allow for the formation of labeled probe hybridization complexes only in the presence of the target. Stringency can be controlled by varying the step parameters as thermodynamic variables. The parameters of the step include, but are not limited to, temperature, alpha (tetra) concentration, salt concentration, chaotropic salt concentration, pH, organic solvent concentration, and the like. These parameters can also be used to control non-specific binding as described in U.S. Patent No. 1, '697. Therefore, certain steps need to be performed under higher stringency conditions to reduce non-specific binding. The reactions described herein can be accomplished in a variety of ways. The reaction components can be added simultaneously or sequentially in a different order, preferably as described below. In addition, the reaction can include a variety of other reagents. Reagents include salts, buffers, neutral proteins (e.g., albumin), detergents, and the like, which can be used to promote optimal hybridization and lateral and/or reduce non-specific or background interactions. Depending on the preparation method and the purity of the stem, 'reagents which improve the efficiency of the assay in other ways, such as protease inhibitors, nuclease inhibitors, antimicrobial agents and the like, may also be used as appropriate. The test data is analyzed to determine changes in the amount of expression of individual genes and the amount of expression between the various states, thereby forming a gene expression distribution. 0 Biological Activity Correlation Assay The present invention provides a compound which modulates the activity of a cancer-associated gene or protein of the present invention. A method of identification or screening. The methods comprise adding a test compound as defined above to a cell comprising a cancer protein of the invention. These cells contain a recombinant nucleic acid encoding a cancer protein of the invention. In another embodiment, a library of candidate agents is tested against a plurality of cells. In one aspect, in physiological signals (eg hormones, antibodies, peptides, antigens, cytokines, growth factors [potential potentials, pharmacological agents (packages: chemotherapeutics), radiation, carcinogens or other cells (ie cells) 'Cell contact υ presence or absence. The assay is evaluated under or prior or subsequent exposure. In another example, the assay is performed at different stages of the cell cycle process. In this way, the person regulating the gene or protein of the present invention is identified. Pharmacologically active compounds are capable of enhancing or interfering with the activity of the cancer 1 protein of the present invention. (4) Other structures are evaluated to identify key structural features of the compound 142769.doc-147-201021828. Providing a method of modulating (e.g., inhibiting) cancer cell division; the method comprising administering cancer (IV). In another embodiment, providing a method of modulating (e.g., inhibiting) cancer; the method comprising administering a cancer modulator In another embodiment, a method of treating a cell or an individual having cancer; the method comprising administering a cancer modulator. ◎ In an embodiment Providing a method of modulating the detachment of cells expressing the basis of the invention. As used herein, the state encompasses parameters of the art such as cell growth, proliferation, 4 activity, function, Cell death, aging, localization, enzymatic activity, signal transduction. In one embodiment, the 'cancer inhibitor is an antibody as described above. In another embodiment the cancer inhibitor is an antisense molecule. The various cell growth, proliferation and metastasis assays are known to those skilled in the art. 9 Qualitative adjustment of the Qualcomm 1 screening to identify appropriate modulators for high-throughput screening. Detection of enhancement or inhibition of cancer gene transcription, inhibition or enhancement of polypeptide expression, and inhibition or enhancement of polypeptide activity. In the examples, the modulators evaluated by high-throughput screening are proteins, usually natural. a fragment of a protein or a naturally occurring protein. Thus, for example, a protein-containing cell extract 4 or a random or directed digest of a protein cell extract is used. The library is screened in the method of the invention. Particularly preferred in this embodiment are libraries of bacterial, fungal, viral and mammalian proteins, the latter of which are preferred and human proteins are particularly preferred. Test compound needle 142769.doc •148- 201021828 The protein class (such as the substrate of the enzyme) or ligand and receptor to which the standard belongs. Use soft agar growth and colony formation method to identify and characterize the regulator. The matrix is attached and grows. When the cell is transformed, it loses this phenotype and grows out of the matrix. For example, the transformed cells can be grown in stirred suspension culture or suspended in semi-solid medium (such as half In solid or soft agar). Transformed cells, when transfected with a tumor suppressor gene, can regenerate normal phenotypes and again require a solid matrix to attach and grow. Soft agar growth or colony formation in assays is used to identify modulators of cancer sequences that, when expressed in host cells, inhibit abnormal cell proliferation and transformation. The modulator reduces or eliminates the growth ability of host cells suspended in a solid or semi-solid medium such as agar. The techniques used for suspension growth or colony formation in suspension assays are described in Freshney, Culture of Animal Cells a Manual of Basic

Technique (3rd edition, 1994). See also Garkavtsev et al. (1996), ibid. ^ Assessing Contact Inhibition and Growth Density Limits to Remember and Characterize Regulators Normal cells are typically grown in cell culture in a flat and organized manner until they contact other cells. When the cells are in contact with each other, they are inhibited by contact and stop growing. However, the transformed cells are not inhibited by contact and continue to grow to a high density with a messy focus. Therefore, the transformed cells grow to a higher density than the corresponding normal cells. This is morphologically detected by forming a disordered monolayer of cells or focal cells. Alternatively, the growth density limit is quantified using a marker that utilizes the balance of '(3Η)-thymidine, and similarly, the Alamar blue assay reveals cell proliferation capacity and 142769.doc • 149· 201021828 The ability of tincture to affect it. See Freshney (1994), ibid. When transformed cells are transfected with a tumor suppressor gene, normal phenotypes can be regenerated and grown to a lower density by contact inhibition. In this assay, the use of a label of saturated density (3H)-thymidine is a preferred method for measuring the growth mobility limit. The transformed host cells can be transfected with cancer-associated sequences and grown at saturation density for 24 hours under non-limiting media conditions. The percentage of cells labeled with (3h)-thymidine was determined by pooled cpm. Contact-independent growth is used to identify modulators of cancer sequences that have caused abnormal cell proliferation and transformation. The modulator reduces or eliminates contact-independent growth and restores the cells to normal phenotype. Growth factors or serum dependence are assessed to distinguish and characterize cells that regulate the transformation of the thank-through to have a serum dependency that is lower than their normal counterpart (see, eg, Temin, J. Natl. Cancer Inst. 37: 167-175 (1966) ;

Eagle et al., j Εχρ Med 131:836 879 (197〇)); , ibid.). This is in part due to the fact that the transformed cells release a variety of growth factors or the degree of serum dependence of the primary cells transformed with the growth factor A can be compared to the control. For example, growth factors or serum dependence of cells are monitored in each method to identify and characterize compounds that modulate the cancer-associated list of the invention. The amount of certain factors that regulate tumor cell release (hereinafter referred to as "tumor-specific markers") is more abundant than that of normal counterparts using the tumor-specific marker content. For example, human gliomas release a higher level of plasminogen activity factor 142769.doc -150- 201021828 (PA) compared to normal sputum cells (see eg Gullino, Angiogenesis, Tumor Vascularization, and Potential). Interference with Tumor Growth, in Biological Responses in Cancer, pp. 178-184 (Mihich (ed.) 1985). Similarly, tumor cells release tumor angiogenic factor (TAF) at levels higher than their normal counterparts. See, for example, Folkman, Angiogenesis and Cancer, Sem. Cancer Biol. (1992), while bFGF is released from endothelial tumors (Ensoli, B et al). A variety of techniques for measuring the release of these factors are described in Freshney (1994), ® Ibid. See also Unkless et al, J. Biol. Chem. 249: 4295-4305 (1974); Strickland & Beers, J. Biol. Chem. 251:5694-5702 (1976); Whur et al., Br. J. Cancer 42:305 312 (1980);

Gullino, Angiogenesis, Tumor Vascularization, and Potential Interference with Tumor Growth, in Biological Responses in Cancer, pp. 178-184 (Mihich (ed.) 1985); Freshney, Anticancer Res. 5:1 11-130 (1985). For example, tumor specific marker levels are monitored in each method to identify and characterize compounds that modulate the cancer associated sequences of the invention. The degree of invasive invasion into the matrigel or extracellular matrix component within the Matrigel used to identify and characterize the modulator can be used as an assay to identify and characterize compounds that modulate cancer-associated sequences. Tumor cells are positively correlated between malignant disease and the invasiveness of cells to matrigel or some other extracellular matrix component. In this assay, tumorigenic cells are commonly used as host cells. The expression of tumor suppressor genes in such host cells reduces the invasiveness of the host cells. The techniques described in the following documents can be used: Canper Res. 142769.doc -151 - 201021828 1999; 59:6010; Freshney (1994), supra. Briefly, the degree of invasion of host cells is measured by using a filter coated with Matrigel or some other extracellular matrix component. Infiltrating into the gel or penetrating to the far side of the filter is rated as invasive 'and histologically depending on the number of cells and the distance traveled by δ, or by pre-labeling the cells with 125 并 and calculating the filter distally Or the bottom of the dish. The radioactivity of sputum is flat. See, for example, Fr e shney (1984), supra. Evaluation of live tumor growth to identify and characterize modulators The effects of cancer-associated sequences on cell growth were tested in transgenic or immunosuppressive organisms. The genetically modified organic system is prepared in a variety of ways that have been accepted by this technology. For example, a gene knockout gene organism is formed, such as a mammal, such as a mouse, in which a cancer gene is split or a cancer gene is inserted. The gene knockout transgenic mouse is formed by inserting a gene or other heterologous gene into the endogenous cancer gene locus of the mouse genome via homologous recombination. The mice can also be formed by mutating the endogenous cancer gene with a mutant form of the cancer gene or by mutating the endogenous cancer gene by exposure to a carcinogen. To prepare a transgenic chimeric animal (e. g., a mouse), the DNA construct can be introduced into the nucleus of the embryonic stem cell. Cells containing newly engineered genetic lesions are injected into the host mouse embryo, which is then implanted into the female recipient. Some of these embryos develop into chimeric mice with germ cells, some of which are derived from mutant cell lines. Therefore, it is possible to obtain a novel mouse strain containing the introduced genetic lesion by breeding a chimeric mouse (see, for example, Capecchi et al., Science 244: 1288 (1989)). Chimeric mice can be obtained according to the following documents: U.S. Patent No. 142, 769. doc-152-201021828, 6, 365, 797, issued on Apr. 2, 2002; U.S. Patent No. 6,107,540 issued on August 22, 2000; Hogan et al., Manipulating the Mouse Embryo: A Laboratory Manual, Cold Spring Harbor Laboratory (1988); and Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, edited by Robertson, IRL Press, Washington, DC, (1987). Alternatively, a variety of immunosuppressive or immunodeficient host animals can be used. For example, genetically athymic "naked" mice (see, eg, Giovanella et al, ® J. Natl. Cancer Inst. 52: 921 (1974)), SCID mice, resected thymic mice, or irradiated mice (see For example, Bradley et al, Br. J. Cancer 38: 263 (1978); Selby et al, Br. J. Cancer 41: 52 (1980)) can be used as a host. Transplantable tumor cells (usually about 106 cells) injected into an isogenic host produce invasive tumors at a high rate, whereas normal cells like the source do not. Cells expressing a cancer-associated sequence are injected subcutaneously or orthotopically into a host that forms an invasive tumor. The mice are then grouped, including the control group and the treatment experimental group (eg, treated with a modulator). After appropriate ❹ time (preferably 4-8 weeks), tumor growth was measured (e.g., by volume or according to its two largest sizes, or body weight) and compared to the control group. A statistically significant reduction (using a Student's T test, for example) can be considered to have been inhibited. Live Hip Identification to Identify and Characterize Regulators The assay to identify compounds with modulatory activity can be performed in vitro. For example, the cancer polypeptide is first contacted with a potential modulator and incubated for a suitable period of time, for example, from 0.5 to 48 hours. In one embodiment, the cancer polypeptide content is determined in vitro by measuring the amount of protein 142769.doc • 153-201021828 or mRNA. The protein content is measured using an immunoassay (such as Western blotting, ELISA, and the like) via an antibody that selectively binds to the cancer polypeptide or a fragment thereof. For the amount of mRNA, amplification using, for example, PCR, LCR or hybridization assays (e.g., Northern hybridization, ribonuclease protection, and dot blotting) is preferred. As described herein, the amount of protein or mRNA is detected using directly or indirectly labeled detection agents such as fluorescent or radiolabeled nuclear, radioactive or enzymatically labeled antibodies and analog. Alternatively, a cancer protein promoter design operably linked to a reporter gene (such as luciferase, green fluorescent protein, CAT or P_gal) can be used. The reporter construct is typically transfected into the cell. After treatment with a potential modulator, the amount of transcription, translation or activity of the gene is reported according to standard techniques known to those skilled in the art (1) palpitations, ibid;

Gonzalez, J. & Negulescu, P. Curr. 〇pin. Biotechnol. 1998· 9:624). ❹ Examined in vitro screening of $# individual genes and gene products. That is, the regulators that express the performance of the two soils themselves are screened by the specific differential expression genes that are required to be separated from their fields by the bells. In the Si: Example, the modulator of the expression of a specific gene is screened for D, 嘥 only one gene or several genes. In another example, the sieve is designed as a bead. Then evaluate ^ "No discovery, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .doc -154- 201021828 Binding assay for identifying and characterizing modulators In the binding assays of the invention, purified or isolated gene products of the invention are generally used. For example, antibodies are produced against the proteins of the invention, and Immunoassay to determine the amount and/or position of the protein. Or 'in the assay, a cell comprising a cancer protein is used. Thus, the methods comprise: combining a cancer protein of the invention with a candidate compound (such as a ligand); and the compound The determination is carried out in combination with the cancer protein of the present invention. Preferred embodiments use human cancer proteins; animal models of human diseases can also be developed and used. Furthermore, other similar mammals can be used as known to those skilled in the art. Proteins. Further, in some embodiments, variants or derivatives of cancer proteins are used.

In general, the cancer protein or ligand of the present invention binds non-proliferatively to insoluble proppants. The support can be, for example, a support having a quarantined sample receiving area (microtiter plate, array, etc.). The insoluble support can be made of any composition that can be combined with the compositions, easily separated from the soluble material, and additionally Compatible with the overall screening method. The surface of the material (4) may be solid or porous and may be in any convenient shape. Examples of suitable insoluble branches (4) include microtiter plates, arrays, membranes and beads. These alternatives are typically made of glass, plastic (e.g., polystyrene), polysaccharide, nylon (nyl〇n), stone sinusoidal fiber, or TeflonTM. Microtiter plates and arrays are particularly convenient due to the ability to perform multiple assays simultaneously with a small amount of reagents and samples. The particular manner in which the composition is combined with the support is not critical as long as it is compatible with the reagents and overall methods of the invention, maintains the activity of the composition 142769.doc -155-201021828 and does not diffuse 'better combination method includes use when An antibody that does not block the ligand binding site or the activation sequence when the protein is attached to the support; directly binds to "viscous" or ionic support; chemically crosslinks; synthesizes proteins or agents on the surface. The protein or ligand/binding agent is bound to the support and then the excess unbound material is removed by washing. The sample receiving area can then be blocked by incubation with bovine serum albumin (BSA), casein or other harmless proteins or other parts. Once the cancer protein of the invention binds to the support, the test compound is added to the assay. Alternatively, the candidate binding agent is bound to the support and then the cancer protein of the invention is added. The binding agent includes a specific antibody, a non-naturally-burning binding agent screened by a screening chemical library, a peptide analog, and the like. Particular attention is paid to the identification of agents that have low toxicity to human cells. A variety of assays can be used for this purpose, including proliferation assays, cAMp assays, labeled in vitro protein-protein binding assays, electrophoretic mobility shift assays, protein binding immunoassays, functional assays (phosphorylation assays, etc.) and the like. Verification. The binding of the test compound (ligand, binding agent, modulator, etc.) to the cancer protein of the present invention can be determined in a variety of ways. The test compound can be labeled and the binding assayed directly as follows: for example, attaching all or part of the cancer protein of the invention to a solid support; adding a labeled candidate compound (eg, a fluorescent label); washing away excess reagent; Determine if the label is present on the solid support. Multiple blocking and washing steps can be used as appropriate. In some embodiments, only one of the components is labeled, for example, the protein or ligand of the invention is labeled. Alternatively, more than one component may be labeled with a different 142769.doc -156-201021828 (for example, for the protein m. It may also be used as a camping light group). The progressively competitive binding is adjusted to set the transfer reagent. In the '::::, the fine is determined by the competitive combination test through the "competitive agent": the hair"; A competitor is a binding moiety that binds to a stem molecule (e.g., a body X peptide: a disease protein). Competing agents include compounds such as anti-[human]: & ligates, ligands and the like. In some cases, competitive binding between the beta and the competitor allows the test compound to be displaced: in the -example the test compound is labeled. Test compound, competing agent, or both are added to the protein for a sufficient time to allow for the knot. . Incubate at temperatures that favor optimal activity (usually between generations and thieves). The incubation period is typically optimized to facilitate, for example, rapid high throughput screening, usually between time and time. Excess reagents are generally removed or washed away. The second component is then added and the presence or absence of the labeled component is followed to indicate binding.一 In one embodiment, a competitor is first added followed by a test compound. Competitor replacement indicates that the test compound binds to the cancer protein and is thereby capable of binding and potentially modulating the activity of the cancer protein. In this embodiment, any of the components can be labeled. Thus, for example, if the competitor is labeled' then the presence of a label in the solution after washing of the test compound indicates replacement of the compound being tested. Alternatively, if the test compound is labeled, the presence of a standard on the support indicates that the substitution has occurred. In an alternate embodiment, the test compound is first added while culturing and washing, followed by the addition of a competitor. The absence of a competitor binding indicates that the test compound 142769.doc • 157-201021828 binds to the cancer protein with a higher affinity than the competitor. Thus, if the test compound is labeled, the presence of a label on the support together with the absence of a competitor indicates that the test compound binds and thereby potentially modulates the cancer protein of the present invention. Therefore, the 'competitive binding method' includes a differential screening method for an agent capable of regulating the activity of the cancer protein of the present invention. In this embodiment, the methods comprise combining a cancer protein with a competitor in a first sample. The second m compound, cancer protein and competitor. For the two types: the combination of 敎 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲 甲The agent then binds to the cancer protein. , or 'Use differential screening to identify candidate drugs that can bind = by: a protein. For example, cancer: A drug that is designed to interact with each other in a rational drug design that does not normally bind to the protein. In addition, the products affecting the natural cancer egg L are also memorized by screening the candidate for the (4) M sex. The ability to enhance or reduce the activity of the 4 protein can be used in assays with positive and negative controls. Preferably, the present invention is performed in at least triplicate to obtain a statistically significant result sample culture sufficient to allow the agent to bind to the protein result. After all the samples, the non-specific binders in the sample were interposed: The amount of the general marker agent that is cultivated. For example, the combination of the test and the right use of the radioactive label, 142769.doc -158- 201021828 can be used to count the sample to determine the amount of compound bound. Most & includes a variety of other reagents. Such agents include agents for promoting protein binding and/or reducing non-specific or background interactions: reagents such as salts, neutral proteins (eg, albumin), detergents, and other agents that modify the efficiency of the shirt. Such as protein transduction: agents, nuclease inhibitors, antimicrobial agents, and the like. A mixture of the components is added in the order in which they are to be combined.

Downregulating or inhibiting the protein of the present invention using a polynuclear material, as described in WO 91/G4753, can be inserted into a cell containing a standard dry lipo acid sequence by forming a conjugate with a ligand binding molecule. Inside. Suitable ligand binding molecules include, but are not limited to, cell surface receptor growth cytokines or other ligands that bind to cell surface receptors. The ligation of the ligand binding molecule preferably does not substantially interfere with the ability of the ligand binding molecule to bind its corresponding molecule or receptor, or to block access to the sense or antisense nucleotide or its binding pattern into the cell. Alternatively, a cancer modulator of a cancer can be introduced into a cell containing a target nucleic acid sequence, for example, by forming a polynucleotide-lipid complex as described in WO 90/10448. It should be understood that the use of antisense molecules or base-off and knock-in models other than for therapeutic methods can also be used in screening assays as described above. Inhibitory and antisense nucleotides In certain embodiments, by using an antisense polynucleotide or an inhibitory small nuclear RNA (snRNA) (ie, with an encoding nucleic acid sequence (eg, a cancer protein, mRNA of the invention) A nucleic acid which is complementary and preferably hybridizable to it, or a subsequence thereof, downregulates or completely inhibits the activity of a cancer associated protein. Anti-142769.doc •159· 201021828 The agglutination of nucleotides and mRNA is reduced by ,, and the translation and/or stability of mRNA is reduced. In the context of the present invention, an antisense polynucleotide may comprise a naturally occurring nucleating acid or a synthetic material formed from a naturally occurring subunit or an ortholog thereof. The antisense polynuclear acid may also have an altered sugar moiety or an interactive sugar linkage wherein the saccharide moiety or the interactive sugar linkage is thiophyllin and other sulfur-containing materials known in the art. The present invention encompasses an analog as long as it functions effectively to hybridize to the nucleotide acid of the present invention. See for example Ws

Pharmaceuticals, Carlsbad, CA; Sequitor, Inc., Natick, MA 0 These antisense polynucleotides can be readily synthesized recombinantly or can be synthesized in vitro. Equipment for this synthesis is sold by several suppliers, including Applied Bi〇Systems. The preparation of other oligonucleotides, such as phosphorothioates and alkylated derivatives, is also well known to those skilled in the art. Antisense molecules as used herein include antisense or sense nucleotides. A sense oligonucleotide can be used to block transcription, for example by binding an antisense strand. Antisense and sense oligonucleotides comprise a single-stranded nucleic acid sequence (rna or DNA) capable of binding to an mRNA (sense) or DNA (antisense) sequence of a cancer molecule. Antisense or sense oligonucleotides of the invention comprise fragments which generally have at least about 12 nucleotides, preferably from about 12 to 30 nucleotides. The ability to obtain antisense or sense oligonucleotides based on cDNA sequences encoding a given protein is described, for example, in Stein & Cohen (Cancer Res. 48:2659 (1988)) and van der

Krol et al. (BioTechniques 6:958 (1988)). Ribozymes 142769.doc -160 - 201021828 In addition to antisense polynucleotides, ribozymes can also be used to target and inhibit the transcription of cancer-associated nucleotide sequences. Ribozymes are RNA molecules that catalyze the division of other RNA molecules. Different types of ribozymes have been described, including class I ribozymes, hammerhead ribozymes, hairpin ribozymes, ribonuclease P, and axe ribozymes. (To review the characteristics of different ribozymes in general, see, for example, Castanotto et al., Adv In Pharmacology 25: 289-317 (1994)) ° The general characteristics of the nuclease are described, for example, in Hampel et al, Nucl. Acids Res. 18:299-304 (1990); European Patent Publication No. 0360257®; U.S. Patent No. 5,254,678. Methods of preparation are well known to those skilled in the art (see, for example, WO 94/26877; Ojwang et al, Proc. Natl. Acad. Sci. USA 90: 6340-6344 (1993); Yamada et al, Human Gene Therapy 1: 39-45 (1994); Leavitt et al., Proc. Natl. Acad Sci. USA 92: 699-703 (1995); Leavitt et al., Human Gene Therapy 5: 11 5 1 -120 (1994); and Yamada et al. , Virology 205: 121-126 (1994)). Use of Modulators in Phenotypic Screening In one embodiment, test compounds are administered to a population of cancer cells having associated cancer performance profiles. As used herein, "administering" or "contacting" means that the modulator is added to the cell in a manner that allows the modulator to act on the cell, whether by ingestion and intracellular action, or by acting on the cell surface. . In some embodiments, a nucleic acid encoding a proteinaceous agent (ie, a peptide) is placed into a viral construct, such as an adenovirus or retroviral construct, and added to the cell to effect expression of the peptide agent, eg, PCT US 97/01019. A tunable gene therapy system can also be used. Applying the regulator to the fine 142769.doc -161 - 201021828

Taichung said that the transcription of the target protein does not have to change. The performance of the piano is flat. The regulator inhibition function will be used as a surrogate marker. As described above, screening is performed to assess genes or gene products. That is, the modulator of the expression of the gene or the gene product itself is screened by identifying a specific differential expression gene that is important in a particular state. Use of Modulators to Affect Peptides of the Invention A variety of assays are used to measure cancer polypeptide activity or cancer phenotype. For example, the effect of a modulator on the function of a cancer polypeptide is measured by examining the above parameters. The effect of the test compound on the polypeptide of the invention is assessed using physiological changes affecting the activity. When using intact cells or animals to determine functional outcomes, multiple effects can be assessed, such as cancer associated with solid tumors, tumor growth, tumor metastasis, neovascularization, hormone release, known genetic markers, and uncharacterized inheritance. Transcriptional changes in markers (eg, by northern blotting), changes in cellular metabolism (such as cell growth or pH changes), and changes in intracellular second messengers (such as cGNIP). A method for characterizing cancer-related sequences 142769.doc -162- 201021828

The performance of multiple gene sequences is associated with cancer. Thus, a condition can be determined based on a mutant or variant cancer gene. In one embodiment, the invention provides a method of fusing a cell comprising a variant cancer gene, e.g., determining whether all or a portion of the sequence of at least one endogenous cancer gene is present in the cell. This can be accomplished using a variety of sequencing techniques. The invention encompasses methods for identifying a cancer genotype in an individual, e.g., determining all or part of a sequence of at least one gene of the invention in an individual. The squatting is generally performed in at least one organization of the individual (e.g., the organization listed in Table π), and may include evaluating different tissues or different samples of the same tissue. The method can include comparing the sequence of the sequenced gene to the sequence of a known cancer gene (i.e., a wild type gene) to determine the presence or absence of a family member, homologue, mutant or variant. All or part of the sequence of the gene can then be compared to the sequence of a known cancer gene to determine if there is any difference. This can be performed using a variety of known homology programs, such as BLAST, Bestfit, and the like. As described herein, the sequence difference between a patient's cancer gene and a known cancer gene is associated with a predisposition to a disease state or disease state. In a preferred embodiment, the cancer gene line is used as a probe for determining the number of cancer gene copies in the genome. The cancer gene can be used as a probe for determining the localization of the chromosome of a cancer gene. When chromosomal abnormalities (such as shifts and similar abnormalities) are identified in cancer loci, information such as chromosomal localization can be used to provide a diagnosis or prognosis. XIII.) Therapeutic Use of RNAi and Small Interfering RNA (siRNA) The present invention also relates to siRNA oligonucleotides, particularly comprising at least a fragment of the 58P1D12 coding region or the 5" UTR region or complement or pair 581> A double stranded Rna of any antisense oligonucleotide of 142769.doc -163- 201021828. In one embodiment, the oligonucleotides are used to clarify the function of 58P1D12 or to modulate or evaluate modulators of 58P1D12 function or performance. In another embodiment, the gene expression of 5 8 to 1012 can be reduced by using 3丨1^8 transfection and the proliferative ability of the transformed cancer cells causing the endogenous antigen is significantly reduced; Measurements of viability metabolic readings showed that cells treated with specific 58P1D12 siRNA showed reduced survival, which was associated with decreased proliferative capacity. Thus the '58P1D12 siRNA composition comprises siRNA (double stranded RNA) to the nucleic acid ORF sequence of the deer 58P1D12 protein or a subsequence thereof; the subsequences generally have 5, 6, 7, 8, 9, 1 , n, 12 , 13, 14, 15, 16, 17, 18 '19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more than 35 The contiguous RNA nucleotide is of a length and contains a sequence that is complementary and non-complementary to at least a portion of the mRNA coding sequence. In a preferred embodiment, the subsequence has 19 - 25 core 4 acid lengths, preferably 2 1 - 2 3 nucleotide acid lengths. RNA interference is a novel method for silencing genes in vitro and in vivo, so small double-stranded RNA (siRNA) is an important therapeutic agent. The ability of siRNA to silence specific gene activity has now been used in animal disease models and is also used in humans. For example, the hydrodynamic injection of a siRNA solution containing siRNA against a specific target into a mouse has proven to be therapeutically effective.

Song's prior work indicates that a class of fully natural nucleic acid small interfering RNA (siRNA) is used as a therapeutic agent, even without further chemical modification (Song, E. et al. "RNA interference targeting Fas protects mice from fulminant hepatitis" Nat· Med. 9(3): 347-51 142769.doc -164- 201021828 (2003)). This work provides the first in vivo evidence that the infusion of siRNA into an animal can alleviate the disease. In this case, the authors will be designed to inject mice into silencing siRNAs that silence the Fas protein, a cell death receptor that induces hepatocytes and other cell death when overactivated during an inflammatory response. The next day, antibodies specific for Fas were administered to the animals. Control mice died of acute liver failure within a few days, while more than 8% of siRNA-treated mice were not seriously ill and survived. The liver cells are about 8〇% to 9%% combined with naked nucleotides. In addition, after 3 weeks, the RNA molecule worked and lost its effect after 10 days. For human therapy, siRNA is delivered using an efficient system that induces long-lasting RNAi activity. A major limitation for clinical use is the delivery of siRNA to appropriate cells. Hepatocytes appear to be particularly susceptible to receiving foreign RNA. Today, because the liver is an organ that is susceptible to nucleic acid molecules and viral vectors, the attractive target is in the liver. However, other tissue and organ targets are also preferred. The use of siRNAs and formulations that promote compounds that cross cell membranes to improve the administration of siRNA during treatment. The chemically modified synthetic siRNA is another serum which is resistant to nucleases and which has a concomitant extension of the duration of action of RNAi. Thus ' siRNA technology treats human malignant diseases by delivering siRNA molecules directed against 58P1D12 to individuals with cancer, such as those listed in Table 1. This siRNA administration slows the growth of cancer cells expressing 58 piD12 and provides anti-tumor therapy to reduce the morbidity and/or mortality associated with malignant diseases. When decomposed in vitro or in vivo, the breakdown of the gene product is clearly 142769.doc -165- 201021828 effective. When an in vitro method is used to detect a decrease in the performance of the 58P1D12 protein, the in vitro efficacy is readily demonstrated by the cells added to the culture (as described above) or applied to an aliquot of a cancer patient biopsy. Get proof. XIV·) sets/products

Kits for use in the laboratory, prognostic, prophylactic, diagnostic, and therapeutic applications described herein are within the scope of the present invention. The kits may comprise: a carrier, a package, or a container partitioned to receive one or more containers (such as vials, test tubes, and the like), each container containing a separate component G to be used in the method.

One and a label or insert containing instructions for use, such as those described herein. By way of example, the container can include probes that are detectable or can be labeled. The probe may be an antibody or a polynucleus specific for the protein or gene or message of the present invention. If the method uses a nucleic acid hybridization method to measure the nucleic acid, the kit may also have a nucleic acid sequence for amplification. A container for nucleotides. The kit may comprise a reporter comprising a reporter (such as a biotin-binding protein, such as avidin or avidin phantom) in combination with a reporter molecule such as an enzymatic label, a fluorescent label or a radioisotope label; this reporter may Combinations such as nucleic acids or antibodies can be used. The kit can include all or part of the amino acid sequence of Figure 2, Figure 2 or Figure 3 or an analog thereof or a nucleic acid molecule encoding such amino acid sequences. The group typically comprises the above-described containers and - or a plurality of other containers associated therewith - which contain the materials required by the commercial and user's stand - including buffers, diluents, filters, needles, syringes; / or instructions for use of carriers, seals, containers, vials and / or test tubes labeled I42769.doc -166 - 201021828; and package inserts with instructions for use. Labels may be present on or along with the container to indicate The composition is used for = therapy or non-therapeutic applications, such as prognosis, prevention, prance or laboratory response, and may also indicate in vivo or in vitro instructions, Their instructions for use as described herein. Inserts or labels that are present on or included with the kit may also include instructions and/or other information. The iron may be located on or attached to the container. When a letter, number or other character is formed or etched into the container itself, the label can be located on the container; when the label is present in a receptacle or carrier that also houses the container, it can be attached to the container, such as a package An inset. The label may indicate that the composition is used to diagnose, treat, prevent, or prognose a condition, such as neoplasia of a tissue listed in the table. The terms "set" and "article" are used synonymously. In an embodiment, an article comprising a composition, such as an amino acid sequence, a small molecule, a nucleic acid sequence, and/or a body, for example, for diagnosis, prognosis, prevention, and/or treatment, such as Table Φ, is provided. A substance formed by a tumor of a tissue, etc. The article typically comprises at least one container and at least one label. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers can be made from a variety of materials. Formed with glass, metal or plastic. The container can hold an amino acid sequence, a small molecule, a nucleic acid sequence, a cell population and/or an antibody. In one embodiment, the container contains a polynucleus for examining the distribution of mRNA expression of the cells. Glycosylates and reagents for this purpose. In another embodiment, the container comprises antibodies for assessing protein expression of 5 8P1D12 in cells and tissues or for related laboratory, prognostic, diagnostic, prophylactic and therapeutic purposes a binding fragment or specific binding egg 142769.doc • 167. 201021828 white; instructions and/or instructions for such use may be included on or in conjunction with the container, reagents and other combinations for such purposes The article or tool may also be included on or in conjunction with the container. In another embodiment, the container contains materials and associated indications and/or instructions for inducing a cellular or humoral immune response. In another embodiment, the container comprises a substance for adoptive immunotherapy, such as cytotoxic T cells (CTL) or helper T cells (HTL), and related instructions and/or instructions; may also include for this purpose Reagents and other compositions or tools. The container may alternatively contain a composition effective to treat, diagnose, prognose or prevent the condition and may have a sterile access port (e.g., the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic needle). The active agent in the composition may be an antibody capable of specifically binding to 58P1D12 and modulating the function of 58P1D12. The article of manufacture may further comprise a second container comprising a pharmaceutically acceptable buffer such as phosphate buffered saline, Ringer's solution, and/or dextrose solution. It may further include other materials required by commercial and user standpoints, including other buffers, diluents, filters, blenders, needles, syringes, and/or package inserts with instructions and/or instructions for use. EXAMPLES Various aspects of the invention are further described and illustrated by the following examples, which are not intended to limit the scope of the invention. Example 1 58P1D12 antigen 142769.doc -168- 201021828 The novel 58P1D12 gene sequence was discovered using the suppression subtractive hybridization (SSH) method known in the art. The 427 bp 58P1D12 SSH sequence was identified from the LAPC xenograft SSH experiment using standard methods. The full-length cDNA pure line of 58P1D12 was isolated from the LAPC-9 AD library. The cDNA (pure line 2) is 25 50 bp in length and encodes 273 amino acid 〇rf (see figure). 58 piDi2 v.l exhibits 100% homology to human cartilage agglutinin. See the following documents for further reference. U.S. Patent No. 7,087,718 (Blu Lu 6118}, 111 〇., 831 ^ Monica, CA) and U.S. Patent Publications us 2〇〇5/〇l36435 (Agensys, Inc., Santa Monica) , CA) 剪58P1D12 splicing variant 馥 splicing variant is a mature ❿ variant produced by the same gene by alternative transcription or alternative splicing. Alternative transcripts are transcripts that originate from the same gene but initiate transcription at different points. Splice variants are mRNA variants of the same transcript that are spliced differently. Hugu > 3 a 丄 共 共. In eukaryotic cells, when genomic DNA is transcribed from the genomic DNA, the initial RNA is spliced to produce a functional mRNA, which is only a right resting. m m is used and is used to translate into amino acid sequences. Thus, a given gene can have a variety of alternative transcripts. Each transcript of JL can have a variety of splice variants. Each transcript variant has a unique exon structure relative to the original transcript, /V 。 . . . has different coding moieties and/or non-coding (5, or 3, end) moieties. A transcript variant of 77 may encode a protein that is similar or similar in function to a similar or similar function, or a protein that encodes and functions, and may be simultaneously expressed in the same U regardless of the same tissue, or may not be Behaved at the same time in the same tissue at 142769.doc 201021828, or at different times in different tissues. Proteins encoded by transcript variants may have similar or different cellular or extracellular localization, such as secretory localization Relative to intracellular localization. Transcript variants are identified by a variety of methods accepted in the art. For example, alternative transcripts and splicing are identified by full-length colonization experiments or by using full-length transcripts and EST sequences. Variants. First, all human ESTs are classified into clusters that are directly or indirectly consistent with each other. Second, ESTs in the same cluster are further classified into subgroups. Synthesize consensus sequences. Compare the original gene sequence to a consensus sequence or other full-length sequence. Each consensus sequence is a potential splice variant of the gene (see, for example, the URL www.doubletwist.com/products/c 1 l_agentsOverview.jhtml). When identifying variants that are not full-length pure lines, the other part of the variant is still well suited for the production of antigens and further selection of full-length splice variants using techniques known in the art. Furthermore, genome-based sequences based on this technique can be utilized. Computer program for identifying transcript variants. Genome-based transcript variant identification programs include FgenesH (A. Salamov and V. Solovyev, "Ab initio gene finding in Drosophila genomic DNA," Genome Research. April 2000; 10 ( 4): 516-22); Grail (URL compbio.ornl.gov/ Grail-bin/EmptyGrailForm) and GenScan (URL genes.mit.edu/ GENSCAN.html). For a general discussion of the identification of splice variants, see, for example, Southan, C., A genomic perspective on human proteases, FEBS Lett. June 8, 2001; 498(2-3): 214-8; de Souza, SJ et al., Identification of human chromosome 22 142769.doc -170- 201021828 transcribed sequences with ORF expressed sequence tags, Proc. Natl Acad Sci US A. November 7, 2000; 97(23): 12690- to further demonstrate transcription Parameters of the variants can utilize a variety of techniques in the art, such as full-length colonization, proteomic validation, PCR-based validation, and 5' RACE validation (see, for example, Proteomic Validation: Brennan, SO et al, Albumin banks Peninsula: a new termination variant characterized by electrospray mass spectrometry, Biochem Biophys Acta. August 17, 1999; 1433(1-2): 321-6; Ferranti P et al., Differential splicing of

Pre-messenger RNA produces multiple forms of mature caprine alpha(sl)-casein, Eur J Biochem. October 1997 1 曰; 249(1): 1-7. For PCR-based validation: Wellmann S et al, Specific reverse transcription-PCR quantification of vascular endothelial growth factor (VEGF) splice variants by LightCycler technology, Clin Chem. 2001 April; 47(4): 654-60; Jia, Η·Ρ. et al., Discoveryofnewhumanbeta-defensins using a genomics-based approach, Gene. January 24, 2001; 263(1-2)·. 211-8. For PCR-based validation and 5' RACE validation: Brigle, KE et al, Organization of the murine reduced folate carrier gene and identification of variant splice forms, Biochem Biophys Acta. August 7, 1997; 1353(2): 191- 8). Genomic regions are known in the art to be regulated in cancer. Alternative transcripts or splice variants of the gene are also regulated when the genomic region of gene 142769.doc -171 - 201021828 is regulated in a particular cancer. It is disclosed herein that 58piDi2 has a specific distribution of expression associated with cancer. The alternative transcripts of 58P1D12 and male variants are also associated with cancers of the same or different tissues and are therefore used as tumor-associated markers/antigens. Four full-length genes and EST sequences were used to identify four transcript variants, designated 58P1D12 ν.2, ν.3, ν·4, and ν·5 (Fig. 1B-Fig. 1E). Example 3 Single Nucleotide Polymorphism of 58P1D12 The single nucleotide polymorphism (SNP) is a single base pair variant at a particular position in the nucleotide sequence. At any given point in the genome, there are four possible base pairs of nucleotides: A/T, C/G, G/C, and Τ/Α. A genotype refers to a particular base pair sequence at one or more positions in an individual's genome. A pure type refers to a sequence of base pairs at more than one position on the same DNA molecule (or the same chromosome of a higher organism), usually in the context of a gene or in the context of several closely linked genes. The Snp present on the cDNA is called cSNP. This cSNP can alter the amino acid of the protein encoded by the gene and thereby alter the protein function. Among individuals, some SNp cause disease transmission; some SNPs contribute to phenotypic variability and response to environmental factors, including diet and drugs. Thus, a combination of SNPs and/or alleles (called phenotypes) has a variety of applications, including diagnosing genetic diseases, determining drug response and dosage, identifying genes responsible for disease, and analyzing genetic relationships between individuals (P. Nowotny , JM Kwon and AM Goate, "SNP analysis to dissect human traits, j Curr. Opin. Neurobiol. 142769.doc -172- 201021828 October 2001; 11(5): 637-641; M. Pirmohamed and BK Park, "Genetic susceptibility to adverse drug reactions," Trends Pharmacol. Sci. June 2001; 22(6): 298-305; JH Riley, CJ Allan, E. Lai and A. Roses, "The use of single nucleotide polymorphisms in The isolation of common disease genes," Pharmacogenomics. February 2000; 1(1): 39-47; R. Judson, JC Stephens and A. Windemuth, "The predictive power of haplotypes in clinical response," _ Pharmacogenomics. 2000 February; 1(1): 1 5-26). SNPs are identified by a variety of methods accepted by this technique (P. Bean, "The promising voyage of SNP target discovery," Am. Clin. Lab. 2001-November 2001; 20(9): 18-20; KM Weiss, "In search of human variation," Genome Res. July 1998; 8(7): 691-697; Μ. M. She, "Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies ," Clin. Chem. February 2001; 47(2): 164-1 72). For example, SNPs can be identified by sequencing a DNA fragment that exhibits polymorphism using gel-based methods such as Restriction Fragment Length Polymorphism (RFLP) and Denaturing Gradient Gel Electrophoresis (DGGE). SNPs can also be discovered by direct sequencing of DNA samples from different individual combinations or by comparing sequences from different DNA samples. With the rapid accumulation of sequence data in public and private databases, SNPs can be found by comparing sequences using a computer program (Z. Gu, L. Hillier and PY Kwok, "Single nucleotide polymorphism hunting in cyberspace," Hum. Mutat. 1998; 142769.doc -173- 201021828 12(4):221-225). SNPs can be tested and individual genotypes or simplex can be determined by a variety of methods including direct sequencing and high-throughput microarray (PY Kwok, "Methods for genotyping single nucleotide polymorphisms," Annu· Rev. Genomics Hum. Genet. 2001; 2:235-258; M. Kokoris, K. Dix, K. Moynihan, J. Mathis, B. Erwin, P. Grass, B. Hines and A. Duesterhoeft, "High-throughput SNP genotyping With the Masscode system," Mol· Diagn·December 2000; 5(4): 329-340) ° Ten SNPs of the initial transcript 58PID 12 vl (Fig. 1A) were identified at the following positions using the above method: 1764 (A/C), 1987 (G/A), 2045 (A/G), 2066 (T/C), 2134 (T/A), 2350 (G/T), 2435 (G/T), 3 02 (G/T), 3 04 (G/T) and 153 3 (C/T). The transcripts or proteins with alternative alleles were designated as variants 58P1D12 v.6 to ν.15 (Fig. 1F). Example 4 Recombination of 58P1D12 in Prokaryotic Systems In order to express recombinant 58P1D12 and 58P1D12 variants in prokaryotic cells, the full-length or partial length 58P1D12 and 58P1D12 variant CDNA sequences were cloned into any of various expression vectors known in the art. Among them. The following one or more regions of the 58P1D12 variant are subjected to the expression: the full length sequence shown in Figure 1, or any of the 8, P, D, 58 P1D12 variants or analogs thereof, 8, 9, 10, 11, 12, 13, 14 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more than contiguous amino acids. 142769.doc •174· 201021828 A. In vitro transcription and translation architecture:

pCRII: a pCRII construct that encodes all or a fragment of the 58P1D12 cDNA for the production of a 58P1D12 sense and antisense RNA probe for RNA in situ studies (Invitrogen, Carlsbad CA). The pCRII vector has a Sp6 flanked by the inserted sequence. And the T7 promoter to drive transcription of the 58P1D12 RNA used as a probe in RNA in situ hybridization experiments. These probes were used to analyze the cell and tissue performance of 58P1D12 at the RNA level. The transcribed 58P1D12 RNA representing the cDNA amino acid coding region of the 58P1D12 gene is used in the Living ® external translation system (such as the TnTTM coupled reticulocyte lysate system (TnTTM)

Coupled Reticulolysate System) (Promega, Corp., Madison, WI)) to synthesize 58P1D12 protein. Β·Bacterial construct: ρΘΕΧ construct: a recombinant 5 8P1D12 protein that is fused to a glutathione S_transferase (GST) protein in bacteria, and all or part of the 58P1D12 cDNA protein coding sequence is cloned into the pGEX family. The GST fusion vector (Amersham Pharmacia Biotech, Piscataway, ❹ NJ). These constructs allow for a controlled expression of the recombinant 58P1D12 protein sequence fused to GST at the amino terminus and fused to the six histidine epitopes (6X His) at the carboxy terminus. The GST and 6X His markers allow for the purification of recombinant fusion proteins from the induced bacteria via a suitable affinity matrix and permit recognition of the fusion protein via anti-GST and anti-His antibodies. The 6X His tag is generated by adding six histidine codons to a selection primer located, for example, at the 3' end of the open reading frame (ORF). A proteolytic cleavage site (such as the PreScissionTM recognition site in PGEX-6P-1) can be used to 142769.doc -175-201021828 which allows GST tagging and 58PID 12 associated protein cleavage. The ampicillin resistance gene and the pBR322 origin allow selection and maintenance of pGEX plastids in E. coli. pMAL construct: a recombinant 58P1D12 protein that is fused to maltose binding protein (MBP) in bacteria, and all or part of the 58P1D12 cDNA protein coding sequence is cloned into pMAL-c2X and pMAL-p2X vectors (New England B' Iolabs, Beverly, ΜΑ) is inherently fused to the MBP gene. These constructs allow for the controllable expression of the recombinant 58P1D12 protein © sequence fused at the amino terminus to MBP and fused at the carboxy terminus to the 6X His epitope tag. The MBP and 6X His markers allow purification of the recombinant protein from the induced bacteria via a suitable affinity matrix and allow recognition of the fusion protein via anti-MBP and anti-His antibodies. The 6X His epitope tag was generated by adding six histidine codons to the 3' selection primer. The factor Xa recognition site allows the pMAL marker to split with 58P1D12. The pMAL-c2X and pMAL-p2X vectors were optimized to express recombinant proteins in the cytoplasm or periplasm, respectively. Periplasmic performance enhances the folding of proteins with disulfide bonds. 〇 pET construct: In order to express 58P1D12 in bacterial cells, all or part of the 58P1D12 w cDNA protein coding sequence was cloned into the vector of the pET family (Novagen, Madison, WI). These vectors allow for the recombination of 58P1D12 proteins with proteins that enhance solubility (such as NusA and thioredoxin; Trx) and epitope markers that facilitate purification and detection of recombinant proteins (such as 6X His and S-). TagTM) is tightly controlled in bacteria with and without fusion. For example, the construct is prepared using the pET NusA fusion system 43.1 such that the 142769.doc -176-201021828 region of the 58P1D12 protein is expressed as an amine-based end fusion with NusA. The cDNA encoding the amino acid 22-213 of 58PlD12 was cloned into the pET-21b vector, expressed and purified from bacteria. Recombinant proteins are used to produce rabbit polyclonal antibodies. C. Yeast construct: pESC construct: in order to express 58P1D12 in the yeast species S. cerevisiae cereWhae) to produce recombinant protein and perform functional studies, all or part of the 58P1D12 cDNA protein coding sequence is selected into the vector of pESC family, each The vector contains one of four selectable markers HIS3, ® TRP1, LEU2 and URA3 (Stratagene, La Jolla, CA). Such vectors allow for the controllable expression of identical plastids with up to two different genes or sequences, containing FlagTM or Myc epitope markers, in the same yeast cell. This system is suitable for demonstrating the protein-protein interaction of 58P1D12. In addition, expression in yeast produces similar post-translational modifications, such as glycosylation and acidification that are present when expressed in eukaryotic cells. pESP construct: In order to express 58P1D12 in the yeast species S. cerevisiae (iSace/mrow), all or part of the 58P1D12 cDNA protein coding sequence was cloned into the vector of the pESP family. These vectors allow for a controlled, high degree of expression of the 58P1D12 protein sequence at the amine terminus or at the carboxy terminus that is fused to the GST that facilitates purification of the recombinant protein. The FlagTK^^ original determinant marker allows for the detection of recombinant proteins via the anti-FlagTN^ body. Example 5 Preparation of Recombinant 58P1D12 in Higher Eukaryotic Systems A. Mammalian Constructs: 142769.doc -177- 201021828 To characterize recombinant 58 P1D12 in eukaryotic cells, the full-length or partial-length 58P1D12 cDNA sequence was cloned into this technology. Any of a variety of performance vectors known in the art. One or more of the following regions of 58P1D12 are rendered in the constructs: amino acids 1 to 273 of 58P1D12 vl or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino acids; v. 2 amino acids 1 to 232; amines v.4 Any of 8, 9, 10, u, 12, 13, 14, 15, 16, 17, 18, 19, 19, 20, 21, 22, 23, 24 of the base acid 1 to 236; or 58P1D12 variant or analogue thereof 25, 26, 27, 28, 29, 30 or more ® bis-amino acids. The construct can be transfected into any of a variety of mammalian cells, such as 293T cells. Transfected 293T cell lysates can be probed with anti-58P1D12 multiple strains of serum as described herein. pcDNA4/HisMax construct: To express 58P1D12 in mammalian cells, the 58P1D12 ORF of 58P1D12 or a portion thereof was cloned into type A pcDNA4/HisMax (Invitrogen, Carlsbad, CA). Protein expression is driven by the cellular giant virus (CMV) promoter and the SP16 translational enhancer. The recombinant protein has XpressTM fused to the amine end and six histidine acids (6X His). The pcDNA4/HisMax vector also contains bovine growth hormone (BGH) polyadenylation signal and transcription termination sequences to enhance mRNA stability, as well as S V40 for free replication and simple vector rescue in cell lines expressing large T antigens. starting point. The Zeocin resistance gene allows selection of the protein in the milk animal cell and the ampicillin resistance gene and the ColEl origin allow for selection and maintenance of the plastid in the large intestine 142769.doc-178-201021828. pcDNA3.1/MycHis construct: To express 58P1D12 in mammalian cells, the 58P1D12 ORF or its portion of 58P1D12 with a consensus Kozak translation initiation site was cloned into type A pcDNA3.1/MycHis (Invitrogen, Carlsbad, In CA). Protein expression is driven by the cellular giant virus (CMV) promoter. The recombinant protein has a myc epitope fused to the carboxy terminus and a 6X His epitope. The pcDNA3.1/MycHis vector also contains bovine growth hormone (BGH) ® polyadenylation signal and transcription termination sequences to enhance mRNA stability, as well as for free replication and simple vector rescue in cell lines expressing large T antigens The starting point of the SV40. The neomycin resistance gene can be used because it allows selection of mammalian cells expressing the protein, and the ampicillin resistance gene and the ColEl origin allow selection and maintenance of the plastid in E. coli. The complete ORF of 58P1D12 ν·1 was cloned into the pcDNA3.1/MycHis construct to generate 58P1D12.pcDNA3.1/MycHis. pcDNA3.1/CT-GFP_TOPO construct: e is 58P1D12 in mammalian cells and allows the detection of recombinant proteins using fluorescence. The 58P1D12 ORF or part thereof with a consensus Kozak translation initiation site is cloned into pcDNA3. 1/CT-GFP-TOPO (Invitrogen, CA). Protein expression is driven by the cellular giant virus (CMV) promoter. The recombinant protein has a green fluorescent protein (GFP) fused to the carboxy terminus, which facilitates non-invasive in vivo detection and cell biology studies. The pcDNA3.1CT-GFP-TOPO vector also contains bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability I42769.doc -179- 201021828, and cell lines containing large tau antigens The SV40 starting point for free replication and simple vector rescue. The neomycin resistance gene allows selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColEl origin allow selection and maintenance of plastids in E. coli. Other constructs fused to GFP at the amine terminus spanning the entire length of the 58P1D12 protein were formed in pcDNA3.1/NT-GFP-TOPO. PAPtag: The 58P1D12 ORF or a portion thereof was cloned into pAPt ag-5 (GenHunter Corp. Nashville, TN). This construct produces an alkaline phosphatase fusion at the carboxyl terminus of the 58P1D12 protein while fused the IgGK signal sequence to the amine terminus. A constitutage body in which an alkaline phosphatase having an amino terminal IgGK signal sequence is fused to an amine terminal of the 58P1D12 protein is also produced. The resulting recombinant 58P1D12 protein is optimized for secretion into the culture medium of the transfected mammalian cells and can be used to identify proteins, such as ligands or receptors, that interact with the 58P1D12 protein. The protein expression is driven by the CMV promoter and the recombinant protein also contains rnyc and 6X His antigen determinants fused at the ruthenium end, which facilitates detection and purification. The Zeocin® resistance gene present in the vector allows selection of mammalian cells expressing the recombinant protein and the ampicillin resistance gene allows selection of plastids in E. coli. pTag5: The 58P1D12 ORF or a portion thereof was cloned into pTag-5. This vector is similar to pAPtag but without alkaline phosphatase fusion. This construct disk produces a 58P1D12 protein with an IgGK signal sequence at the amino terminus and a myc and 6X His epitope at the carboxy terminus for efficacious and affinity purification 142769.doc -180· 201021828. The resulting recombinant 58P1D12 protein is optimized for secretion into the culture medium of the transfected mammalian cells and used as an immunogen or ligand to identify proteins (such as ligands or receptors) that interact with the 58P1D12 protein. Protein expression is driven by the CMV promoter. The Ze〇cin resistance gene present in the vector allows selection of mammalian cells expressing the protein and the ampicillin resistance gene allows selection of plastids in E. coli.

PsecFc :

The 58P1D12 ORF or a portion thereof was cloned into pSecpc*. The psecFc vector was assembled by cloning human immunoglobulin Gl (IgG) Fc (hinge, CH2, CH3 region) into pSecTag! (Invitrogen, California). This construct produced an IgG1 Fc fusion at the carboxy terminus of the 58P1D12 protein, and at the same time fused the IgGK signal sequence to the end. A 58P1D12 fusion using the murine IgG1 Fc region can also be used. The resulting recombinant 58 piD12 protein is optimized for secretion into the culture medium of the transfected mammalian cells and can be used as an immunogen or for identifying proteins (such as ligands or receptors) that interact with the 58P1D12 protein. Protein expression is driven via the CMV promoter. The hygromyCin resistance gene present in the vector allows selection of mammalian cells expressing the recombinant protein and the ampicillin resistance gene allows selection of plastids in E. coli. pSRa construct: To produce a mammalian cell line that is constitutively expressing 58P1D12, the 58P1D12 ORF or a part thereof is selected into a construct. Double 142769.doc -181 - 201021828 tropism by transfecting the pSRa construct into the 293T_10A1 encapsulated cell line or co-transfecting pSRa with the helper plastid (containing the deletional encapsulation sequence) into 293 cells And an ecotropic retrovirus. Infection of a variety of mammalian cell lines with retroviruses results in the integration of the selected gene, 58P1D12, into the host cell line. Protein expression is driven by long terminal repeats (LTR). The neomycin resistance gene present in the vector allows selection of mammalian cells expressing the protein and the ampicillin resistance gene and the ColEl origin allow selection and maintenance of the plastid in E. coli. The retroviral vector can then be used to infect and produce a variety of cell lines, such as P C3, NIH 3T3, TsuPrl, 293 or rat-Ι cells. Additional pSRa constructs are formed that fuse the epitope tag (such as the FLAGTM tag) to the carboxy terminus of the 58?1012 sequence to allow for detection using anti-Flag antibodies. For example, the FLAGTM sequence 5' gat tac aag gat gac gac gat aag 3' (SEQ ID NO: 20) is added to the re-priming primer at the 3' end of the ORF. Other pSRa constructs were formed to generate the myc/6X His fusion protein of the amine-terminal GFP and the terminal GFP and the full-length 58P1D12 protein. Other viral vectors: Form other constructs for viral-mediated delivery and expression of 58P1D12. High viral titers that cause high expression of 58P1D12 are achieved in viral delivery systems, such as adenoviral vectors and herpes amplicon variants. The 58P1D12 coding sequence or a fragment thereof was amplified by PCR and subcultured into the AdEasy trans-t vector (Stratagene). Recombination and viral encapsulation were performed according to the manufacturer's instructions to generate an adenoviral vector. Alternatively, the 58P1D12 coding sequence or a fragment thereof is cloned into an HS V-1 vector (Imgenex) to produce a sore viral vector. The viral vector is then used to infect a variety of cell lines, such as PC3, NIH 3T3, 293 or rat-1 cells. 142769.doc -182- 201021828 Regulatory Expression System: To control the expression of 58P1D12 in mammalian cells, the coding sequence of 58P1D12 or a portion thereof is selected for regulation in mammalian expression systems, such as the T-Rex system (Invitrogen), GeneSwitch System (invitr〇gen) and tightly regulated serotonin (Sratagene). These systems allow for the study of the transient and concentration dependent effects of recombinant 58P1D12. These vectors are then used to control the performance of 58P1D12 in a variety of cell lines (such as pc), NIH 3T3, 293 or rat-sputum cells. B. Baculovirus expression system is the production of recombinant 58P1D12 protein in the baculovirus expression system, and the 58P1D12 ORF or a part thereof is selected into the baculovirus transfer vector pBlueBac 4.5 (InVitr〇gen), and the transfer vector is at the end of 1 Provide the ms_ tag. Specifically, PBlUeBaC-58PlD12 was co-transfected with the helper plastid pBac_N_Blue (Invitrogen) into SF9 (Grassland worm/r sinus) insect cells to produce recombinant baculovirus (see mvitrogen instructions for details). The baculovirus is then collected from the cell supernatant and purified by plaque assay. The recombinant 58ρ1Ε12 protein was then produced by infecting HighFive insect cells (Invhrogen) with purified baculovirus. The recombinant 58piD12 protein can be detected using an anti-58P1D12 or anti-His-labeled antibody. The 58P1D12 protein can be purified and used in a variety of cell-based assays or as an immunogen to produce polyclonal antibodies and monoclonal antibodies specific for D12. Example 6 142769.doc 183- 201021828 Antigenic distribution and distribution of primary structure amino acids, such as hydrophilicity (ΗοΡΡ τ·ρ., woods KR·, 1981. P roc. Natl. Acad. Sci. USA 78:3824-3828 Hydrophobicity (Kyte J-, Doolittle RF, 1982. J. Mol. Biol. 157: 105-132); percentage of reachable residues (Janin J., 1979 Nature 277: 491-492); average flexibility (Bhaskaran R., & P〇nnuswamy P.K., 1988. Int. J. Pept. Protein Res. 32:242-255); β-angle (Deleage, G., R〇ux B. 1 987 Protein Engineering 1 : 289-294); and other properties that may be used in this skill, such as those available on the ProtScale website, to identify the antigenic regions of each of the 58P1D12 and 58P1D12 variant proteins. The above amino acid distributions of the 58P1D12 variant were generated using the following ProtScale analysis parameters: 1) the window size was 9; 2) the center of the window tb 'the window edge weight was 100 °/. And 3) the amino acid distribution values are normalized between 〇 and 1. Hydrophilicity, hydrophobicity, and percentage distribution of reachable residues are used to determine hydrophilic fatty acid fragments (ie, hydrophilicity and percent residue distribution values greater than 〇5 denier hydrophobic distribution values less than 〇.5h. Exposure to an aqueous environment can be present on the surface of proteins, and thus can be used for immunological recognition of antibodies. Immunological recognition by antibodies. Mean flexibility and β-turn distribution determine the ammonium acid fragments that are not bound in the secondary structure (also That is, the β-turn distribution value and the average flexibility distribution value are larger than 〇.5), such as β-sheet and α-helix. These regions are also more positively exposed on the protein and thus can be used for immune recognition, such as immunological recognition by antibodies. The antigenic sequence of the torn DU variant protein indicates that an immunogen can be produced using, for example, the above 1427S9.doc-184-201021828 distribution. The immunogen can be any 5, 6, 7, 8 of the 58P1D12 protein variant listed in Figure 1. 9, 1〇, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50 or more than 50 contiguous amines Acid or a corresponding nucleic acid encoding the same. The peptide immunogen of the present invention may comprise: a peptide region comprising an amino acid position having a hydrophilic distribution value of more than 0.5, at least 5 amino acids in the figure, and an amino acid increment of any integer; including hydrophobicity a region of the amino acid having a distribution value of less than 0.5, a peptide region having at least 5 amino acids of the formula, and an amine acid group increment of any integer; including an amino acid position having a percent residue distribution greater than 0.5 a peptide region having at least one of 5 amino acids in the figure and an amino acid increment of any integer; including an amino acid position having an average flexibility distribution value greater than 0.5, and at least 5 having a pattern Amino acid, amino acid increment is any integer peptide region; and amino acid position including β-turn distribution value greater than 0.5, at least 5 amino acids in the figure, amino acid increments are any integer Peptide immunogens of the invention may also comprise a nucleic acid encoding any of the peptide regions described above. All immunogens (peptides or nucleic acids) of the invention may be embodied in human unit dosage form or may be compatible with human physiology. Pharmaceutical excipient composition The secondary structure of the 58P1D12 protein and the 58P1D12 variant (ie, the alpha helix extension and the predicted presence and location of the random coil) is based on the ΗΝΝ-hierarchical neural network method (NPS@: Network Protein Sequence Analysis TIBS 2000). Volume 25, Issue 3, March [291]: 147-15〇Combei C., Blanche C., Geourjon C. and Deldage G., https:// I42769.doc -185- 201021828 pbil.ibcp.fr/ Cgi-bin/npsa_automat.pi?page=npsa_nn.html) » The ExPasy Molecular Biology Server (https://www.expasy ch/t〇〇Is/) access is predicted by its primary amino acid sequence. Analysis showed that the 5 8p J D12 variant 1 consisted of 24.18% alpha helix, 18.68% extended strand and 57.140/. Random curl composition. 58P1D12 variant 2 consisted of 19.83% alpha helix, 18.97% extended strand and 01.21% random coil. 581>1 dying 12 variant 3 consists of 322〇%〇1 helix, 15.25% extended temple and 52.54% random coil. The possible presence of the transmembrane domain in the 58P1D12 and 58P1D12 variant proteins was analyzed using a variety of cross-flag prediction algorithms from the ExPasy Molecular Biology Server on the World Wide Web. Example 7 Production of 58P1D12 Multiple Muscles of Resistance Multiple antibodies can be produced in a mammal, for example by one or more injections of an immunizing agent and an adjuvant as needed. The immunizing agent and/or adjuvant is usually injected into the mammal by multiple subcutaneous or intraperitoneal injections. In addition to immunization with full-length 58p1Dl2 protein or 58P1D12 variant, computer algorithms can also be used to design immunogens based on kine acid sequence analysis that contain immunogenic features that are recognized by the immune system of the immunized host (see heading Examples of "antigenic distribution and secondary structure" are predicted to have hydrophilic, flexible, beta corner configurations and to be exposed on the surface of the protein. For example, a recombinant bacterial fusion protein or peptide containing a hydrophilic, flexible, beta-corner region of a 581>1012 protein variant can be used as an antigen to produce multiple antibodies in New Zealand white rabbits (New Zea丨 and White rabbhs). . For example, in the name 58P1D12 variant, the regions include (but are not limited to 142769.doc • 186 - 201021828) amino acid 19-30, amino acid 49-66, amino acid 70-82, amine The base acid 88-115, the amino acid 131-145, the amino acid 165-203, the amino acid 243-255 and the amino acid 262-273. It is suitable to combine an immunizing agent with a protein known to be immunogenic in an immunized mammal. Examples of such immunogenic proteins include, but are not limited to, keyhole spirulina (KLH), serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. In one embodiment, a peptide encoding the amino acid 102-11 5 of 58P1D12 variant 1 was conjugated to KLH and used to immunize rabbits. Alternatively, the immunizing agent may comprise all or part of a 58 piD12 ® variant protein, an analog thereof or a fusion protein. For example, the 58P1D12 variant 1 amino acid sequence can be used in recombinant DNA techniques with a variety of fusion protein partners well known in the art (such as GSH-S-transferase (GST) and HIS-labeled fusion proteins). Any one of them merges. In another embodiment, the recombinant technology and the pET21b expression vector are used to transform the 58P1D12 variant! The amino acid 22_213 is fused with His. The protein was then visualized, purified and used to immunize 2 rabbits. The fusion proteins are purified from the induced bacteria using the appropriate affinity. Other recombinant bacterial fusion proteins that may be used include maltose binding protein, LacZ, thioredoxin, NusA or immunoglobulin constant regions (see the section entitled "Preparation of 58P1D12 in prokaryotic systems", and Current)

Protocols In Molecular Biology, Volume 2, Module 16, Frederick M. Ausubul et al., 1995; Unsley, PS, Brady, W., Urnes, M., Grosmaire, L., Damle, N., A Ledbetter, J. (1991) J. Exp. Med. 174, 561-566). Protein antigens expressed by mammals 142769.doc -187· 201021828 can also be used in addition to fusion proteins of bacterial origin. Such pits are expressed by mammalian expression vectors (such as Tag5 and Fc fusion vectors) (see the section entitled "Preparation of recombinant 58P1D12 in eukaryotic systems"), retaining post-translational modifications such as glycosylation present in natural proteins. . In one embodiment, the amino acid 22-213 of 58P1D12 variant 1 is cloned into a Tag5 mammalian secretory vector and is expressed in 293T cells. The recombinant protein was purified from the tissue culture supernatant of 293T cells stably expressing the recombinant vector by #metal chelate chromatography. The purified D & § 5 58 卩 1012 protein was then used as an immunogen. It is suitable to mix or emulsify the 1 antigen in an adjuvant which enhances the immune response of the host animal during the immunization schedule. Examples of adjuvants include, but are not limited to, complete Freund's adjuvant (CFA) and MPL-TDM adjuvant (monophosphonyl lipid A-synthesis dicorynomycolate) ° in typical In the protocol, up to 200 pg (usually 100-200 gg) of the fusion protein or peptide mixed with KLH and mixed in complete Freund's adjuvant (CFA) is used to remove the rabbit in a subcutaneous manner. Next, up to 200 pg (usually 100-200 pg) of immunogen in incomplete Freund's laryngeal (IFA) is injected subcutaneously into the rabbit every two weeks. Test blood was collected approximately 7-10 days after each immunization and used to monitor the titer of antiserum by ELISA. To test the reactivity and specificity of immune sera (such as rabbit serum immunized with the His fusion of 58P1D12 variant 1 protein), the full-length 58P1D12 variant 1 cDNA was cloned into the pCDNA 3.1 myc-his expression vector (Invitrogen, see The title is "An example of preparing recombinant 5 8P1D12 in a eukaryotic system". After transfecting the 骒 construct into 293T cells, 142769.doc •188- 201021828 was used to detect cell lysates by anti-58P1D12 serum and anti-His antibody (Santa Cruz Biotechnologies, Santa Cruz, CA) using Western blot technique to determine the pair. Specific reactivity of denatured 58P1D12 protein. In addition, immune sera were tested against 293T and other recombinant 58P1D12 expressing cells by glory microscopy, flow cytometry, and immunoprecipitation to determine the specificity of native proteins. Western blotting, immunoprecipitation, fluorescence microscopy, and flow cytometry can also be performed using endogenous cells expressing 5 8 P1D12 to test for reactivity and specificity. Purifying the antibody reactive with the fusion partner sequence by passing it through an affinity column containing the fusion partner or in the context of an unrelated fusion protein', thereby purifying from the 5 8p 1 Antisera from rabbits immunized with 〇12 variant fusion proteins such as GST and MBP fusion proteins. For example, an antiserum obtained from a GST_58P1D12 variant 1 fusion protein is first purified by a column covalently coupled to a GST protein and a substrate (BioRad, Hercules, Calif.). φ is affinity-purified by a column consisting of a covalently coupled MBP_58P1D12 fusion protein and an Affigel matrix. The serum is then further purified by protein G affinity chromatography to isolate the IgG lysate. A column matrix composed of immunogen or free peptide to affinity purify serum from other rabbits immunized with His-tagged antigen and peptide, and fusion partners to exclude serum. Example 8 Production of 58P1D12 monoclonal anti-e (MAb) In one embodiment For the treatment of 58P1D12 & 58P1D12 variant 142769.doc -189- 201021828 degree monoclonal antibody ("MAb") contains an antigenic epitope specific for each protein, or an antigenic epitope specific for the common sequence between variants. Individual antibodies that bind, internalize, disrupt or modulate the biological functions of 58piD12 or 58P1D12 variants, such as disruption and ligand, substrate and binding The ligand interactor. The immunogens used to produce the MAbs include immunogens designed to encode or contain: an extracellular domain or an intact 5 8P1D12 protein sequence, a region predicted to contain a functional motif, and a computer via an amino acid sequence The region of the 58 piD12 protein variant with antigenicity was predicted to be analyzed. The immunogen includes peptides and recombinant proteins such as tag5_58P1D12 (a purified ms-marked protein expressed by a mammal) or PET-58P1D12 (a recombinant protein expressed by Escherichia coli). Further, mice were immunized with cells engineered by retroviral transduction to express high levels of 58P1D12 variant 1 (such as RAT158piDi2). To generate a MAb against 58P1D12, firstly, 5-50 pg of protein immunogen or a mixture of 1〇6 and 1〇7 of 58P1D12 expression cells mixed in a suitable adjuvant is used to place the mouse at the foot pad (Fp). Immunity. Examples of suitable adjuvants for sputum immunization are TiterMax (Sigma) for primary sputum injection and alum gel for subsequent immunization. After the initial injection, the mice were immunized twice a week until the appropriate specific potency was observed. After sacrifice, the lymph nodes were removed and their B cells were harvested for power to induce cell fusion. During the immunization process, test blood is collected to monitor the titer and specificity of the immune response. In most cases, once 142769.doc -190-201021828 is properly reacted and specific as determined by ELISA, Western blot, immunoprecipitation, fluorescence microscopy or flow cytometry analysis, Fusion and fusion tumor production were performed using electro-cell fusion (ΒΤΧ, ECM2000). In one embodiment, the invention provides a monoclonal antibody designated Ha8-4c4.1 (which comprises Ha8-4c4.1 VH and Ha8-4c4_l VL pure line 1-B3), referred to herein as Ha8-4c4.1 . The antibodies listed above were shown to react with and bind to cell surface 58P1D12 (as evidenced by flow cytometry) or to bind to fixed 58P1D12 (as evidenced by ELISA). MAbs against 58P1D12 were generated using XenoMouse technology® (Amgen Fremont, Fremont, CA), where the murine heavy and K light chain loci have been inactivated and most of the human heavy and kappa light chain immunoglobulin loci Inserted. After immunizing XenoMice, which produces human gamma, with 58P1D12-pET recombinant protein, a MAb designated Ha8-4c4.1 was produced. 58P1D12 MAb (Ha8_4c4.1) specifically binds to cells expressing recombinant 58P1D12 and is endogenous cell surface 58P1D12 expressed in cancer xenograft cells. The fusion tumor producing the antibody named Ha8-4c4.1 was delivered to the American Type Culture Collection (ATCC) on August 5, 2008 (via Federal Express), mailbox 1549, Manassas, VA 20108, and designated Deposit number PTA-9404. After isolation of mRNA from the corresponding fused tumor cells using Trizol reagent (Life Technologies, Gibco BRL), the DNA coding sequence of 58P1D12 MAb Ha8-4c4.1 was determined. Total RNA was purified and quantified. Use Gibco BRL Superscript 142769.doc -191- 201021828

The Preamplification system sensitizes the first strand of cDNA from total RNA via oligo (dT) 12-18. The first strand of cDNA was amplified using a human immunoglobulin variable heavy chain primer and a human immunoglobulin variable light chain primer. The PCR product was cloned into the pCRScript vector (Stratagene). The pure line is sequenced and the heavy chain variable region and the light chain variable region are determined. Nucleic acid and amino acid sequences of the heavy chain variable region and the light chain region are shown in Figures 2 and 3. The ratio of the 58P1D12 antibody to the human Ig germline is shown in Figures 4A-4C. Example 9 Screening, Identification, and Characterization 58P1D12 MAb Screening, identification, and characterization using a combination of assays known in the art Antibodies produced by the procedures set forth in the 58P1D12 monoclonal antibody (MAb) examples, including ELISA, FACS, affinity characterization by surface plasmon resonance (BIAcore) ("SPR"), antigen determination Base classification, affinity with recombinant 58P1D12 and 58P1D12 on the cell surface. A. 58P1D12 Human MAb Screening by FACS A primary fusion tumor screening was performed on MAb against 58P1D12 by FACS analysis. The screening protocol was as follows: 50 μl/well of the fusion supernatant (purified) or purified antibody (continuous dilution) was added to a 96-well FACS plate and expressed with 58P1D12 (endogenous or recombinant cells, 50,000) Cell/well) mixed. The mixture was incubated at 4 ° C for two hours. At the end of the incubation, the cells were washed with FACS buffer and at 4. . The cells were incubated with 1 〇〇 μΐ detection antibody (anti-hlgG-PE) for 45 minutes. At the end of the incubation, the cells were washed with FACS buffer 142769.doc -192-201021828, fixed with a nail and analyzed using FACScan. Analyze data using the CellQuest Pro software. Positive fusion tumors identified from the initial screening were transferred to 24-well plates and supernatants were collected for confirmatory screening. Confirmatory screening is performed using FACS analysis and other methods known in the art. B. 58P1D12 Human MAb Screening by ELISA The 58P1D12 MAb was screened by ELISA to determine antibody isotypes. The screening protocol used was as follows: ❹ ELISA plate was coated with Tag5-58P1D12-ECD or anti-hlgG antibody. Array test antibodies were added to the plates and incubated for 1 hour. After washing the disc to wash away unbound antibody, the antibody bound to the antibody was detected by the following HRP ligation: anti-hlgGl, anti-hIgG2, anti-hlgK and anti-hlgL. C. 58P1D12 Human MAb Screening by SPR SPR allows for the identification and immediate characterization of the kinetics and affinity of protein-protein interactions and is therefore a useful technique for selecting and characterizing MAbs for the desired target antigen. SPR analysis was used to screen and characterize the fusion tumor supernatant and purified MAb against 58P1D12. The fusion tumor screening for 58卩1012]^8 13 was performed by SPR biosensor (31 〇〇^ 3 000) as follows: diluted with test buffer (HBS-P, 10 pg/ml BSA) 50 μl/well of fusion tumor supernatant (purified) to 1.5-2 pg/ml was added to a 96-well plate (BIAcore) and via a goat anti-human covalently immobilized on the surface of a CM5 sensor wafer FcY pAb captures MAb (20 μΐ). Three (3) MAb-containing fusion tumor supernatants were tested for each test (cycle) via channels 2, 3 and 4 of the flow cell, with channel 1 remaining as a reference for non-specific binding. In each individual channel, 142769.doc -193- 201021828 before the binding of the antigen to the captured MAb, 60 μL of assay buffer was injected onto the surface of the wafer at a flow rate of 20 μΐ/min for use as the captured MAb. Reference to baseline offset. Sixty microliters (60 μM) of 150 nM purified recombinant 58P1D12 was then injected onto the surface of the wafer at the same flow rate of 20 μΐ/min to measure antigen binding. After each round of antigen binding to the MAb, the surface was regenerated by injection of 100 mM phosphoric acid (for 1 minute) to allow any MAb captured to detach from the surface. Data analysis was performed using BiaEvaluation 4.1 and CLAMP software (Myszka and Morton, 1 998). After deducting the reference and normalizing the reaction to the amount of captured Θ MAb, the data was fully fitted using a 1:ι binding model. Affinity is calculated via the binding rate constant and the dissociation rate constant. As is apparent to those skilled in the art, the slow off rate generally indicates a higher total affinity for MAb. Preliminary affinity data and dissociation rates were used as the basis for selection criteria for the therapeutic MAb of 58P1D12. D. Affinity determination by FACS The binding affinity of Ha8-4c4.1 to 5 8 PID 12 (i.e., 3T3-〇 58P1D12) expressed on 3T3 cells was tested. Briefly, fifteen (15) samples of the purified Ha8-4c4.1 MAb diluted 1:2 at 4 ° C at a final concentration of 80 nM to 0·0049 nM with 3T3-58P1 d 12 cells ( Cultivate overnight with 50,000 cells per well. At the end of the incubation, the cells were washed and incubated with anti-hlgG-PE detection antibody for 45 minutes at 4 °C. After washing the unbound detection antibody, the cells were analyzed by FACS. The average fluorescence intensity (MFI) values as listed in Table vi (A) were obtained. Enter the MFI value into Graphpad Prisim software and analyze it with one point combination (hyperbolic) equation of Y=Bmax*X/(Kd+X) to produce Ha8- shown in Table VI(B) with 142769.doc -194- 201021828 4c4 1 4c4.1 and 58P1D12 啬 female aunt b# dip rabbit and curve. Bmax is Ha8-

Ε·Affinity determination by SPR The binding affinity of the group 58P1D12 was tested by SPR (Biac〇re 3000). In short, ® is obtained on the surface of the CM5 sensor wafer. On average, 12 MAb and purified weight 58P1D12 MAb captures about 150 RU of 58P1D12 MAb per round. A series of 5-6 recombinant 58P1D12 dilutions ranging from 1 nM to 200 nM were injected onto this surface to generate binding curves (sensogram) using BiaEvaluation (Biacore, Inc.) or CLAMP software ( Myszka and Morton, 1998) fully fit the binding curve to the 1:1 interaction model. The affinity of the 58P1D12 MAb was determined using the equation KD = kdiss / kassoc, expressed as KD as defined by the dissociation rate constant and the binding rate constant. Affinity data and dissociation rates and affinity analysis by FACS (see Section D above) are part of the selection criteria for the MAb of 58P1D12. Example 10 Anti-Infectious Immune-Mediated Cytotoxicity ADCC (antibody-dependent cellular cytotoxicity) is an immune-mediated lytic cytotoxicity against cells that bind to antibodies that target specific cell surface antigens. The immune cells recognize the Fc portion of the antibody via binding to the Fey receptor on the surface of leukocytes, monocytes, and NK cells, thereby causing lytic erosion leading to cell death of 142769.doc -195·201021828. Briefly, cells engineered to express the target antigen 58P1D12 were incubated with chromium for 1 hour in vitro. After washing with fresh medium, the labeled cells were combined with 2.5 mg/mI of human MAb against 58P1D12 and newly isolated peripheral blood mononuclear cells at different effector-target cell ratios (E:T ratio). Cultivate. At 37. After 4 hours, the cells were gently centrifuged and the supernatant containing 51Cr released from dead cells was counted in a p-counter. The results demonstrate that when the ratio of effector cells to target cells is increased (Ε:τ), antibody-dependent killing of cells is enhanced. Example 11 Generation of F(ab')2 Fragment

The F(ab')2 fragment that produces the MAb can be used to study the role of the 2MAb molecule retaining its bivalent antigen binding site but lacking the immune effect Fc domain in an in vitro and in vivo therapeutic model. The protocol was as follows: MAb HI-1.10 in 2 mg of mg2 mM sodium acetate buffer (pH 4.5) was incubated for a specified time in the presence and absence of immobilized pepsin (Pierce. R〇ckford 111〇.) by Protein A Chromatography removes intact MAb and digested Fc fragments. Observed that undigested intact MAb is not reduced, aliquots of digested material collected at the specified time are not reduced and finally digested F(ab,)2 SDS-PAGE Coomasie stained gel with reduced sample of the product. This reagent can be used to treat animals with tumors exhibiting 58P1D12. The anti-tumor activity observed by this antibody fragment can be used to intrude biological activity. The activity mediated by the immune-dependent mechanism is distinguished. This reagent can also be used to detect 58P1D12 protein in immunohistochemistry, ELISA, and other diagnostic immunizations J42769.doc • 196-201021828. Example 12 Recombinant DNA Methods The human MAb was recombined to express 58P1D12 MAb in transfected cells, and the 5 8P1D12 MAb variable heavy and light chain sequences were separately selected from human heavy chain IgG1 and light chain IgK constant. Upstream of the region, the entire 58P1D12 MAb human heavy and light chain cassette was cloned downstream of the CMV promoter/enhancer via the selection vector. The downstream of the MAb coding sequence includes the polyadenylation site® point. The constructs expressing recombinant 58P1D12 MAb were transfected into 293T, Cos and CHO cells. The binding of 58P1D12 MAb secreted by recombinant cells to cell surface 58P1D12 was evaluated. Example 13 Activity outside 58P1D12 MAb inhibition studies Enhanced migration and invasion It is a hallmark of cancer cell phenotype. Therefore, the 58P1D12 MAb was evaluated in vitro to determine the effects on cell migration and cell invasion. — MAb Ha8-4c4.1 inhibits the migration and invasion of chamber cells using MDCK/58P1D12 cells in the Boyden Transwell chamber. The migration assay evaluates the effect of 58P1D12 MAb Ha8-4c4.1 on cell migration by adding 4xl04 to 0.1% FBS plus 25 pg/mL of control MAb or MAb Ha8-4. 1 of 10 in 4.1 (1; 1 (758 1012 cells were applied to the chamber of 8〇7 (16111^3118, 611 equipment and allowed to migrate 10% of the FBS in the lower chamber for 16 hours to assess migration. Calcein AM dye will be used Capture Cells were labeled for 30 min and photography on the bottom of the filter via the 142769.doc -197- 201021828

The MetaMorph imaging software quantifies the amount of cellular fluorescence (migration). As shown in Figure 5, MAb Ha8-4c4.1 inhibited cell migration by about 45%, while the negative control MAb did not inhibit cell migration (*ρ<〇.〇〇〇ι). In addition, the effect of 58P1D12 MAb Ha8-4c4.1 on tumor cell invasion was evaluated. In this assay, a Boyden Transwell chamber was coated with a layer of Matrigel® for cell invasion. MAb Ha8-4c4.1 or isotype matched MAb (25 pg/mL) was added to 4xl04 OVCAR-5/58P1D12 cells in 0.1% FBS in the upper chamber of the Matrigel®-coated device. These cells were allowed to invade for 10 hours at 10% FBS loaded in the lower chamber. Cells bound to the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. As shown in Figure 6, MAb Ha8-4c4.1 significantly inhibited cell invasion by up to 75°/ compared to control MAb. (*p<〇.〇〇01). Comparison of the active functional activity of 58P1D12 MAb The complete human 58P1D12 MAb Ha8-4c4.1 (YlK), Ηα8-6·1 (γ2κ) and Ha8-7·1 (γ1κ) were tested by tumor cell migration and tumor cell invasion assay. Migration of tumor cells was assessed using the MDCK/58P1D12 cells in a Boyden Transwell chamber migration assay. Apply 4x1 〇4 of CK_1% FBS plus 25 pg/mL of control MAb or MDP/5 8P1D12 cells in 5 8P1D12 MAb to the chamber above the Boyden Transwell device and allow the cells to be in the lower chamber 1%% FBS migrated for 16 hours to assess migration. The cells captured on the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. The amount of cell glory (migration) was quantified via the MetaMorph imaging software. The results indicate that Ha8-4c4.1 and Ha8-7.1 MAb inhibit cell migration' 142769.doc -198- 201021828

The Ha8-6.1 MAb does not inhibit migration. Tumor cell invasion was assessed using a B〇yden Transwell chamber coated with a layer of Makigel® for cell invasion. Briefly, 5 8P1D12 MAb or isotype matched control MAb (25 pg/mL) was added to 4 χ 104 OVCAR-5/58P1D12 cells in 0.1% FBS in a chamber coated with Matrigel®. These cells were allowed to invade for 10 hours at 10% FBS loaded in the lower chamber. Cells bound to the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. ® results indicate that Ha8-4c4.1 and Ha8-6.1 MAb inhibit tumor cell invasion, while Ha8-7.1 MAb does not inhibit invasion. (Figure 7). MAb Ha8-4c4.1 inhibits 58P1D12-induced HUVEC tube formation. The effect of MAb Ha8-4c4.1 on 58P1D12 ECD-induced HUVEC tube formation. Recombinant 58P1D12 ECD (3 gg/mL) was added to HUVEC (5 x 104/well) in 0.1% FBS with 30 pg/mL of the isotype matched control MAb or MAb Ha8-4c4.1. The cells were then applied to Matrigel® and allowed to form tubes for 16 hours. As shown in Figure 8, control MAb did not affect 58P1D12 ECD-induced HUVEC tube formation, while MAb Ha8-4c4.1 inhibited tube formation by 50% (*p=0.005). Taken together, these data demonstrate that MAb Ha8-4c4.1 exhibits potent inhibitory activity against 58P1D12 function in vitro. Their functions (migration and invasion) induced by the expression of 58P1D12 and their function (tube formation) by the addition of ECD proteins are strongly inhibited by MAb. These conclusions support the use of MAb Ha8-4c4.1 in the form of treatment targeting cancers that exhibit 58P1D12. Comparison of 58P1D12 MAb for HUVEC tube formation 142769.doc -199· 201021828 for HUVEC tube formation assay. Complete human 58卩1012]^8 13 1^8-4ο4.1(γ1κ), Ηα8-6.1(γ2κ ) and Ha8-7.1 (YlK). Briefly, recombinant 58P1D12 ECD (3 pg/mL) was added to 30 Pg/mL of 58P1D12 MAb Ha8-4c4.1, Ha8-6.1 or Ha8-6.1 to HUVEC in 0.1% FBS (5xl04/ In the hole). The cells were then applied to Matrigel® and allowed to form tubes for 16 hours. Calculate the number of tubes. The results showed that all three 5 8P1D12 MAbs inhibited tube formation, expressed as (+). (Figure 9). Example 14 Anti-caries-mediated secondary killing 58P1D12 MAb mediates saporin-dependent killing in 3T3-58P1D12 cells. On day 1, 3T3-58P1D12 cells (1000 cells/well) were seeded in 96-well plates. Secondary sputum, will contain 1x concentration of the designated primary antibody together with a 2-fold excess of Sabinin toxin (Advanced

Targeting Systems, San Diego, CA) Jointed against humans (Hum-

An equal volume of medium of Zap) or anti-goat (Gt Ig Sap) polyclonal antibody was added to each well. The cells were incubated for 4 days at 37 degrees Celsius. At the end of the incubation period, Biosource was added to each well and incubation continued for an additional 4 hours. After excitation at 530 nm, the fluorescence emission of three replicate samples at 590 nm was determined. The results in Figure 10 indicate that the 58P1D12 antibody Ha8-4c4.1 mediates saporin-dependent cytotoxicity in 3 3-5 8P1D12 cells, whereas the control non-specific human IgG1 (H3-1.4.1.2) has no effect. These results indicate that a drug or cytotoxic protein can be selectively delivered to cells expressing 3T3-58P1D12 and other 58P1D12 142769.doc-200-201021828 using an appropriate anti-58P1D12 MAb (e.g., Ha8_4c4.1).

Example 15 58P1D12 tumor growth-promoting live beta internal assay The effect of 58P1D12 protein on tumor cell growth was assessed in vivo by assessing tumor formation and growth of 58P1D12 or 58P1D12-deficient cells. For example, a 3Τ3 or ovarian cancer cell line containing tkNeo empty vector or 58P1D12 was subcutaneously injected into each flank of SCID mice. At least two strategies can be used: (1) constitutively expressing 58P1D12 under promoter regulation, such as a self-virus (such as polyomavirus, fowlpox virus (UK 2, 211, 504 published July 5, 1989), a constitutive promoter obtained from the genome of adenovirus (such as adenovirus 2), bovine papilloma, avian sarcoma virus, giant cell virus, retrovirus, hepatitis B virus and monkey virus 4 (SV40), Or a heterologous mammalian promoter such as an actin promoter or an immunoglobulin promoter, the restriction being that the promoter is compatible with the host cell system; and (2) regulating expression under the control of an inducible vector system, Such as ecdysone, tetracycline and the like, the restriction is that the promoters are compatible with the host cell system. The tumor volume is then monitored by caliper measurement when the palpable tumor is present and tracked over time to determine whether the 58P1D12 expressing cells grow at a faster rate and whether the tumor produced by the 58P1D12 expressing cells exhibits invasive changes. (eg increased metastasis, angiogenesis, reduced reactivity to chemotherapeutic drugs, j,). In addition, the same cells can be orthotopically implanted into mice to determine whether 58p丨D 〖2 has an effect on local growth in the peritoneum and whether 58P1D12 affects cell turnover. 142769.doc -201· 201021828 The ability to move (Miki T et al. , Oncol Res. 2001; 12:209; Fu X et al, Int J Cancer. 1991, 49: 938). The effect of 58P1D12 on tumor formation and growth was assessed by intra-articular injection of ovarian or prostate tumor cells. This assay can also be used to determine 58P1D12 inhibition of candidate therapeutic compositions such as 58P1D12 intrabody, 58P1D12 antisense and ribozyme. Example 16 58P1D12 Single Plant Against Inhibition of Intratumoral Tumor Growth The remarkable expression of 58P1D12 on the cell surface of tumor tissues and its restricted expression in normal tissues made 58P1D12 an excellent target for antibody therapy. Similarly, 58P1D12 is a target for T cell-based immunotherapy. Therefore, the therapeutic efficacy of 58P1D12 MAb in a human ovarian cancer xenograft mouse model and a human prostate cancer xenograft mouse model was evaluated. The effects of antibodies on tumor growth and metastasis formation were studied in a mouse cancer xenograft model (eg, subcutaneous, intra-tibial, and intraperitoneal). The antibody can be unconjugated as described in this example, or can be combined with a therapeutic form as understood in the art. Subcutaneous (s.c.) tumors were generated by injecting 5 χ l〇 4-1 〇 6 cancer cells mixed with Matrigel (Collaborative Research) at a 1:1 dilution into the right abdomen of male SCID mice. To test the effect of antibodies on tumor formation, that is, to start injecting antibodies on the day of tumor cell injection. As a control, purified mouse IgG (ICN) or PBS was injected into the mouse, or purified MAb recognizing an unrelated antigen not expressed in human cells was injected into the mouse. In the preliminary study, no difference was found between mouse IgG or PBS for tumor growth. Tumors 142769.doc -202- 201021828 Dimensions were determined by caliper gauge and tumor volume was calculated as length x width x height. Mice with subcutaneous tumors larger than 1.5 cm in diameter were sacrificed for intra-articular injection, and mice were anesthetized with an appropriate amount of ketamine/xylazine/Acepromazine cocktail. A 5 mm incision was made in the prepared left knee area to expose the humerus. The 271/2-type needle is inserted into the tibia through the proximal end of the bone and along the tail. A 10 μΐ pre-formed cell suspension was injected into the space formed by the 271/2-type needle using a Hamilton syringe. The incision was sutured using a 6_ 丝 silk suture. Tumor growth was monitored using a caliper gauge. At the end of the experiment, the animals were sacrificed and the left tibia and right tibia were weighed via an electronic balance. Tumor weight was determined by subtracting the weight of the right tibia with the tumor from the weight of the contralateral tibia without the tumor. Ovarian tumors usually metastasize and grow in the peritoneal cavity. Therefore, the growth of mouse intraperitoneal ovarian tumors was performed by directly injecting two million cells into the peritoneum of female mice. Monitor the general health, physical activity Φ and appearance of the mouse until it becomes dying. At the time of sacrifice, the peritoneal cavity can be examined to determine tumor burden and the lungs harvested to assess whether to metastasize to the distal site. Alternatively, death can be used as the end point. The mice were then isolated for grouping for proper treatment' wherein 58P1D12 or control MAb was injected intraperitoneally (i.p.). The advantage of the xenograft cancer model is the ability to study new blood vessel formation and angiogenesis. Tumor growth depends in part on neovascularization. Although capillary vasculature and the resulting jk fluid network originate from the host, the initiation and construction of the new vasculature is regulated by xenograft tumors (David〇ff et al., Clin Cancer I42769.doc -203- 201021828).

Res. (2001) 7:2870; Solesvik et al., Eur J Cancer Clin Oncol. (1984) 20:1295). The effects of antibodies and small molecules on neovascularization are studied according to procedures known in the art, such as IHC analysis of tumor tissue and its surrounding microenvironment. 58P1D12 MAb inhibits the formation of ovarian and prostate cancer xenografts. The 58P1D12 MAb also delays the growth of established orthotopic tumors and prolongs the survival of tumor-bearing mice. These results indicate that 58P1D12 MAb can be used to treat local and advanced ovarian and prostate cancers and their cancers listed in Table I. 58P1D12 monoclonal antibody: A monoclonal antibody against 58P1D12 was generated as described in the example entitled "Production of 58P1D12 monoclonal antibody (MAb)". MAbs were characterized by the ability of MAb to bind 58P1D12 by ELISA, Western blot, FACS and immunoprecipitation. The epitope on the 58P1D12 protein is recognized by epitope mapping data for 58P1D12 MAb as determined by ELIS A and Western analysis. MAb was purified from ascites or fusion tumor tissue culture supernatant by protein-G or protein-A agarose chromatography, dialyzed against PBS, filter sterilized and stored at -20° (by: £1^) Perform protein assays on 18 or 00 28〇11^1. Prepare a mixture of therapeutic MAbs or mixtures containing individual MAbs and treat subcutaneous and peritoneal membranes that receive 3T3-58P1D12, OVCAR5-58P1D12 or LAPC9-AI tumor xenografts. Intra- or intra-tibia injections of cells. Cell lines and xenografts: 3T3-58P1D12 and OVCAR5-5 8P1D12 cells were maintained at 142769.doc -204- 201021828, supplemented with L-glutamine and 10% FBS In DMEM and RPMI, LAPC9-AI and LAPC9-AD tumor xenografts were relayed to male ICR severe complex immunodeficiency (SCID) mice (Taconic Farms) from 6 weeks to 8 weeks with a subcutaneous needle implant ( Craft, N. et al., Nat Med. 1999, 5: 280). Preparation of a single cell suspension of LAPC9-AI or LAPC9-AD tumor cells as described in Craft et al. Other cell lines were also used. 58P1D12 MAb inhibition Growth of xenograft tumors showing 58P1D12 using 3T3-5 8P1D12, OV The CAR5-58P1D12 and LAPC9-AI Tumor® models tested the effect of 58P1D12 MAb on tumor formation. Compared with the subcutaneous tumor model, the intraperitoneal and intra-tibial models caused local tumor growth, metastasis in the distal site, deterioration of the patient's health and subsequent Death. These features make the intraperitoneal and intra-humeral models more representative of human disease progression and allow us to track the therapeutic effects of 58P1D12 MAb on clinically relevant endpoints. The 58P1D12 MAb or control antibody injections were administered over a 4-week period with established Mice bearing tumors in the intraperitoneal and tibia. The mice in the two groups were allowed to establish a high tumor burden to ensure a high rate of metastasis in the lungs of the mice. The mice were then sacrificed and analyzed by IHC analysis. Whether there are tumor cells in the liver, bone and lung. 58P1D12 antibody inhibits tumor formation and delays the growth of established tumors and prolongs the survival of the treated mice. Therefore, the main clinically relevant endpoint (tumor growth), survival The 5 8P1D12 MAb is effective in terms of prolongation and health. 58P1D12 MAb Ha8-4c4.1 3T3 in SCID mice -58P1D12 Effect of growth of tumor xenografts In this experiment, 3T3-58P1D12 cells (5.〇xl〇6 cells) were embedded in 142769.doc -205 - 201021828 in Matngel and implanted on the third day In the right abdomen of male SCID mice. Mice were randomized into the same sputum (n=10 per group) and treatment was started intraperitoneally using 500 Ha8_4c4.1 or isotype control MAb (twice a week for a total of 8 doses). Tumor growth was monitored using a caliper gauge every 3 to 4 days. As a result, on the 27th day, Ha8_cc4.1 inhibited the growth of the 3T3_58p1D12 tumor xenograft grown in SCID mice by about 78% as compared with the control antibody alone. The difference in tumor volume between control tumors and Ha8-4c4.1 treated tumors was statistically significant (ρ<〇·〇〇〇1) when analyzed using the Mann-Whitney U test. (Figure 11). In another experiment, 3T3-5SP1D12 cells (5.0χ104 fine moon packs) were embedded in Matrigel and surgically implanted into the right tibia of male SCID mice on the third day. Tumors were established for 7 days at which time mice were randomized (n=10 per group). Treatment was initiated intraperitoneally using an I.5 mg loading dose of HA8-4c4.1 or an isotype control MAb followed by 750 pg of each corresponding Mab (twice per week for a total of 6 doses). Tumor growth was monitored using a caliper measurement every 3 to 4 days. The results demonstrated that on day 24, Ha8-4c4.1 inhibited the growth of established 3T3-58P1D12 limb tumor xenografts grown in mouse tibia by approximately 63% (<0.01, compared to treatment with control antibody treatment). Use the Mann-Whitn_ey U test). (Figure 12). Effect of 58P1D12 MAb Ha8-4c4.1 on the growth of LAPC9-AD tumor xenografts in SCID mice In this experiment, the stock solution of LAPC9-AD tumors was enzymatically eliminated, 142769.doc -206- 201021828 On the third day, 1.5 million live cells were subcutaneously implanted into the right tibia of male SCID mice. Mice were randomized into groups at the same time (^ (7) per group) and treatment was initiated intraperitoneally with 500 sighs of Ha8-4c4.1 or isotype control human IgG1. Animals were treated twice a week for a total of 1 dose until the 32nd. At the end of the study, the animals were sacrificed and the left tibia and right tibia were weighed via an electronic balance. Tumor weight was determined by subtracting the weight of the tumor-free right tibia from the weight of the contralateral tibia without tumor. As a result, on the 32nd day, Ha8-4c4.1 inhibited the growth of the mouse tibia t-grown LAPC9-AD prostate cancer xenograft by 60% as compared with the control antibody treatment. The difference between the control tumor weight and the tumor weight of Η&8_4c4.1 was statistically significant (p=0.0057) when using Student's t-test analysis. (Figure 13). Effect of 58P1D12 MAb Ha8_4c4.l on the growth of established nested tumors in mice In this experiment, tumor cells φ (2.〇xl〇6 cells) expressing 〇vcar5-58PlD12 were implanted into female SCIDs. Inside the right tibia of the mouse. On the next day, mice were randomized (n=10 per group) and 500 HaSjM] or isotype pairs were used, and 3⁄4 human IgGl was started intraperitoneally (i.p.). Animals were treated twice a week for a total of 12 doses until 42nd. At the end of the study (p. 42), the animals were sacrificed and the right tibia and left tibia were weighed via an electronic balance. The tumor weight is the measurement obtained after subtracting the weight of the contralateral tibia without tumor, and the tumor weight is plotted as a graph. The results demonstrate that Ha8-4c4.1 can be effective against Ovcar5-58PlD12 tumors in a single agent form compared to control antibody treatment, allowing growth to be inhibited by I42769.doc • 207- 201021828 56% (p=0.0002, using Mann-Whitney U test) ). (Figure 14). In another experiment, 〇vcar5-58PlD12 tumor cells (2·〇χ1 06 cells) were injected into the peritoneum of female SCID mice on day 0. When the tumor was fully established, the mice were randomized (n=i5 per group) and treated with the intraperitoneal approach using 500 Ha8-4c4.1 or the isotype control human igGl. Animals are treated with antibodies twice a week as long as they survive. The health and survival of the mice were monitored and recorded in the number of weeks during the study. The results demonstrated that mice with well established ovarian tumors treated with HA8-4c4.1 survived a median of 69 days and that the control MAb treated mice survived a median of 37 days. The median survival days of mice treated with HA8-4c4.1 increased by 32 days with a sigmoid (p=〇.〇〇66, using the Logrank test). (Figure 15). Effect of combination of 5SP1D12 MAb Ha8-4c4.l and carboplatin on the growth of human prostate cancer xenografts in mice In this experiment, 'Assessed as a monotherapy and a combination with the chemotherapeutic carboplatin Ha8-4c4 .1 The ability to establish an androgen-independent prostate tumor xenograft (LAPC9-AI). The LAPC9-AI tumor stock solution was enzymatically digested, counted, and 1. 5 x 106 cells were surgically implanted into the right tibia of male SCID mice on the following day. The tumor was allowed to establish for 7 days, at which time the animals were randomly divided into four different groups (n==l each). Beginning with stage 7, intraperitoneal administration of a loading dose (2 mg) of Ha8-4c4.1 MAb or an isotype control human IgGl followed by a maintenance dose (ι·〇mg) of the corresponding MAb twice a week for a total of 7 Agent. Carmine (40 mg/kg) was administered intravenously (i.v.) to mice on days 7, 11, 15, 19, 22 and 26. On page 33, all mice were sacrificed at 142769.doc -208-201021828 and the tumor was excised and weighed with an electronic balance. The results demonstrate that Ha8-4c4_1 is very effective in a single agent form and inhibits tumor growth by 76% compared to control antibody treatment (p=〇77). Carboplatin monotherapy also inhibited tumor growth, resulting in a 87% inhibition of tumor growth (p=0.0001). Combination therapy with Ha8-4c4.1 and carboplatin enhanced inhibition and resulted in 97% inhibition of tumor growth compared to control antibody alone (p<0.0001). Statistically significant ® differences were also demonstrated when comparing the tumor weight of the Ha8-4c4.1 plus carboplatin treatment group to the tumor weight of the control MAb plus carboplatin treatment group (P = 0.0243). Statistical analysis was performed initially using the Kruskal-Wallis test to determine significance in each group. Next, a student t test or a Mann-Whitney U test was applied to each pair. (Figure 16). The results of these experiments demonstrate that 58P1D12 MAb can be used for therapeutic and diagnostic purposes to treat and control the cancers listed in Table I. Example 16 Therapeutic and diagnostic use of 58P1D12 anti-caries in humans 10 58P1D12 MAb can be safely and effectively used for diagnostic, prophylactic, prognostic and/or therapeutic purposes in humans, preferably for the treatment of the cancers listed in Table I . Western blotting and immunohistochemical analysis of cancer tissues and cancer xenografts with 58P1D12 MAb demonstrated that the staining was extensive and broad in cancer, but the amount of staining in normal tissues was significantly lower or undetectable. Detection of 58P1D12 in cancer and metastatic disease demonstrates that MAb can be used as a diagnostic and/or prognostic indicator. 5 8P1D12 antibodies are therefore used in diagnostic applications, such as immunohistochemical analysis of ovarian biopsy samples to detect cancer in suspected patients. 142769.doc -209- 201021828 58P1D12 MAb is heterozygously bound to cancerous cells as determined by flow cytometry. Therefore, 58P1D12 MAb is used in diagnostic whole body imaging applications such as radioimmuno scintigraphy and radioimmunotherapy (see for example

Potamianos S. et al., Anticancer Res 2〇 (2乂): 925-948 (2000)), in order to detect local and metastatic cancer presenting 58P1D12. The extracellular domain of 58P1D12 is released or released into the extracellular environment (as seen for the basic linonic acid diesterase B10 (Meerson, N. R., Hepatology 27: 563-568 (1998))), allowing diagnosis with 58P1D12 MAb Post-test 58P1D12 in serum and/or urine samples of suspected patients. The 58P1D12 MAb that specifically binds 58P1D12 is used in therapeutic applications to treat cancers that exhibit 58P1D12. The 58P1D12 MAb is used in an unbound form and in a conjugated form (the which is linked to one of a variety of therapeutic or imaging modalities well known in the art, such as prodrugs, cytotoxic agents, enzymes or immunoisotopes). In preclinical studies, the efficacy of unconjugated 58P1D12 MAb and conjugated 58P1D12 MAb in preventing tumors and inhibiting growth in the SCID+ murine cancer xenograft model was tested. (See, for example, the example entitled "5 8P1D12 monoclonal antibody inhibits growth of viable tumors in vivo"). The 58P1D12 MAb and the uncoupled 58P1D12 MAb were treated as a treatment alone or in combination with other treatment groups as described in the Examples below for use in human clinical trials. Example 17 Human Roadbed Test for Treatment and Diagnosis of Human Carcinoma via S8P1D12 MAb 58P1D12 MAb for use in the treatment of certain tumors according to the present invention, 142769.doc-210-201021828 such as those listed in Table i . Tumors (such as those listed in Table I) are currently preferred indications based on a number of factors, including 58 piDi2 performance. Three clinical approaches have been successfully employed for each of these indications. I.) Combination therapy: In combination therapy, a patient is treated with a 58P1D12 MAb in combination with a chemotherapeutic or anti-neoplastic agent and/or radiation therapy. Treatment of primary cancer targets by adding 58P1D12 MAb to first-line and second-line therapy, such as those listed in Table I, such as reduction in tumor mass and reduction in standard chemotherapy The dose is evaluated and the program design focuses on effectiveness. Such dose reductions allow for other and/or prolonged therapies because of the reduced dose-related toxicity of the chemotherapeutic agent. In several assisted clinical trials, 58P1D12 MAb is used in combination with chemotherapeutic agents and other therapies known in the art. In one embodiment, there is a synergistic effect when treating a tumor (including a human tumor) with a 58P1D12 antibody in combination with a chemotherapeutic agent or radiation or a combination thereof. In other words, the inhibition of tumor growth by the 58P1D12 antibody increased more than expected when combined with a chemotherapeutic agent or radiation or a combination thereof. Synergism can be presented as &apos;e&quot;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos;&apos; Synergism is preferably evidenced by cancer remission, where treatment with either the naked 58P1D12 antibody or the combination of the 58P1D12 antibody and the chemotherapeutic agent or radiation is not expected to be resolved. H.) Monotherapy: For the use of 58P1D12 in a monotherapy of tumors 142769.doc -211 - 201021828 MAb ' can be administered to a patient in the absence of a chemotherapeutic or anti-neoplastic agent. In one embodiment, a monotherapy is performed clinically on a terminal cancer patient with extensive disease metastasis. The patient showed a slight stabilization of the disease. Test the effect of refractory patients with cancerous tumors. HI·) Engagement 58P1D12 MAb: For the treatment of cancers such as ovarian cancer, the 58P1D12 MAb of the present invention can be conjugated to a toxin, such as with a calicheamicin (eg, Myl〇targTM, Wyeth-Ayerst, Madison, NJ, a antagonist Tumor antibiotic echinomycin-conjugated recombinant humanized IgG4 κ antibody) or conjugated with melamine (eg, a taxane-based tumor-activated prodrug 平台ρ platform, 瘳ImmunoGen, Cambridge, ΜΑ, see also, for example, US Patent 5, 4丨6,〇64) or with Ollistatin E (Nat Bi〇techn〇l. July 2003 '21(7): 778-84. (Seattle Genetics)). IV·) Imaging agent: A radiolabeled antibody can be used as a diagnostic and/or imaging agent by binding a radionuclide (e.g., iodine or hydrazine (Im, Y9)) to a 58P1D12 MAb. In this effect, the labeled antibody is localized to a metastatic lesion of a solid tumor and a cell exhibiting 58P1D12. For the 58P1D12 MAb used as an imaging agent, these anti-systems are used as adjuvants for solid tumor hand treatments' for pre-operative screening and post-operative tracking to determine which tumor residuals and/or recurrences. In one embodiment, the (111111)_58?11)12 antibody is used as an imaging agent in a phase 1 human clinical trial in a patient with a cancer of 58P1D12 (see also, for example, Divgi et al. #加/. /« W. 83 _·97_ 104 (1991)) ^Fat patients with standard anterior and posterior gamma cameras. The results showed that primary lesions and metastatic lesions were identified. Dosing and Routes of Administration 142769.doc -212· 201021828 As will be appreciated by those of ordinary skill, dosing considerations can be determined by comparison with similar products of clinical orders. Therefore, an anti-58P1D12 antibody can be administered at a dose ranging from 5 to 400 mg/m4 in a safety study, for example, using an anti-58P1D12 antibody in a lower agent for its target affinity relative to a known antibody. In addition to the parameters used by the skilled artisan to determine similar dosage regimens, anti-58P1D12 antibodies as fully human antibodies have a slower clearance compared to chimeric antibodies; therefore, such fully human anti-58P1D12 The amount of antibody administered to the patient is low, perhaps in the range of 5 〇 to 3 〇〇 mg/m 2 , and is still effective. Compared with the conventional dose measurement in units of 瓜 仏 ' The measure of surface area is a convenient measure of administration for all body types of patients designed to include infants to adults. However, as will be appreciated by those skilled in the art, the desired dosage unit can be used. Two different routes can be used. The method delivers anti-58piDi2 antibody. Conventional intravenous delivery is a standard delivery technique for a variety of tumors. However, for peritoneal tumors, tumors such as nests, bile ducts, other ducts, and the like Intraperitoneal administration has been shown to be beneficial in obtaining high doses of antibodies at the tumor and also minimizing antibody clearance. In a similar manner 'some solid tumors have vascular structures suitable for local perfusion. Local perfusion allows for high presence at tumor sites Dosage of antibodies and minimize short-term clearance of antibodies. Trampoline Development Program (CDP), review: CDP combined with adjuvant therapy, monotherapy, and imaging as an imaging agent to develop anti-58P1D12 antibodies. The trial initially demonstrated safety. And then demonstrate the efficacy of repeated doses. The trial is to compare the standard chemotherapy with the standard 142769.doc • 213· 201021828 Therapy plus the open label of the anti-58P1D12 antibody. As you know, one standard that can be used in patient recruitment is by living Tissue examination of the amount of 58P1D12 expression in the patient's tumor as determined. As with any therapy based on protein or antibody infusion, safety considerations are primarily concerned with: (1) cytokine release syndrome, ie hypotension, fever, trembling, chills; 1) an immunogenic response to the substance (ie, the patient is resistant to Body therapy produces human antibodies, or HAHA reactions and (out) poisoning of normal cells that exhibit 58P1D12. Standard tests and follow-ups are used to monitor each of these safety considerations. Anti-581 &gt;1〇12 antibodies were found to be administered It is safe for human beings. In the whole article of this application, it cites multiple website materials, publications, patent applications and patents. (The website is based on the consistent resource locator of the website. (Uniform Resource Locator) or the URL reference. The disclosure of each of the references is hereby incorporated by reference in its entirety in its entirety herein in The individual aspects of the invention are described, and any functional equivalents are within the scope of the invention. Various modifications of the present invention will be apparent to those skilled in the art in the <RTIgt; Such modifications or other embodiments may be practiced without departing from the true scope and spirit of the invention. Example 18 Designation of Peptide Mapping and Intacting Molecules of the Conjugates of Hosting Number PTA-9404 142769.doc -214· 201021828 Quantitative Analysis Using Peptide Mapping and Amine End Sequencing Techniques and Complete Molecular Weight Analysis for Fusion of Designated Accession Number PTA-9404 The Ha8_4c4.1 monoclonal antibody produced by the tumor was analyzed. Since it is observed that there are two light chain sequences corresponding to Ha8-4c4·1 VL pure line 1-B3 and pure line 2-A7 in the deposited fusion tumor, these studies are intended to prove that the deposit is secreted by the deposit or the like. The identity of the antibody. The amino acid sequence data generated by the experiments indicated that the fusion tumor only secreted a single monoclonal antibody corresponding to Ha8-4c4.1 VL pure line 1-B3. This ® outer molecular weight analysis is consistent with the analysis of the predicted molecular weight of the pure 1-B3. The data generated by these studies led to the conclusion that the deposited fusion tumor only secreted antibodies containing the Ha8-4c4.1 VH heavy chain and the Ha8-4c4.1 VL pure line 1-B3 light chain. Example 19 Recombinant functional analysis of Ha8-4c4.1 VL pure line 1-B3 monoclonal strain expressed in Chinese hamster ovary cells (CHO) was obtained from Ha8-4c4.1 VL pure line 1-B3 (SEQ. ID NO · The amino acid of the light chain of amino acid residues 1 to 134) and the pure line of 2-A7 (amino acid residues 1 to 133 of SEQ. ID NO: 18) and the light chain κ constant The polynuclear ATS of the region is fused and colonized in the expression vector. The expression vector containing the two light chains was transfected into Chinese hamster ovary (CHO) cells. Both chains express light bond proteins in the cell. The 1 - B 3 pure line exhibits a protein with a predicted molecular weight. In contrast, the molecule 1 of the protein expressed by the 2-A7 pure line in the cell exhibited greater than the predicted molecular weight. This result is consistent with the protein retention of the leader sequence. 0 142769.doc -215- 201021828 The 1-B3 and 2-A7 expression vectors were co-transfected with constant and heavy chain sequences to allow assembly of recombinant monoclonal antibodies. When co-transfected with a polynucleotide encoding the heavy chain Ha8-4c4_l VH (containing the sequence shown by amino acid residues 1 to 146 of SEQ. ID NO: 17), only the sequence containing 1-B3 is contained. CHO cells expressing the vector secrete fully assembled monoclonal antibodies. The 2-A7 sequence does not support recombination. The expression of monoclonal antibodies and the above molecular weight data suggest that the 2-A7 light chain has not been treated to remove the leader sequence. If this hypothesis is correct, retaining the 2-A7 light chain of the leader leader may explain the inability to secrete appropriately assembled monoclonal antibodies. The ability of recombinant monoclonal antibodies secreted by CHO cells transfected with the sequences of pure 1-B3 and Ha8-4c4.1 VH to bind to the 58P1D12 protein was tested. The recombinant antibody display binds to the 58P1D12 protein with the same affinity as the secreted monoclonal antibody. These results support the results described in Example 18, indicating that the monoclonal antibodies secreted by the fusionoma of the assigned accession number PTA-9404 contain the pure light chain of the pure line 1-B3. Table Table I: Organizations showing 58P1D12 in malignancy

Table II: Amino acid abbreviation single letter three letter full name F Phe phenylalanine L Leu leucine S Ser serine 142769.doc -216- 201021828 Y Tyr tyrosine c Cys cysteine w Trp tryptamine Acid P Pro Proline H His Histamine Q Gin Glutamine Amine R Arg Ginsic Acid I lie Isoleucine M Met Methionine T Thr Sulfate N Asn Aspartame K Lys Amino Acid V Val proline A Ala alanine D Asp aspartate E Glu glutamic acid G Gly glycine Table III: Amino acid substitution matrix modified by GCG software 9.0 BLOSUM62 amino acid substitution matrix (block substitution matrix) to make. The higher the value, the higher the likelihood of substitution in the relevant natural protein. Q-1 p-1 N-2 MV L-1 K-1 ii T1 - H-2 Go F-2 E-1 D-2 Co A 4 -3 I -3 -3 -2 -4 -3 -3 2 -3 -3-3 -3 -4 -2 -3 R-1-3-2 -3 -3 -3 -3 • A Y-2 W-3 V ο To D -3 -4 -3 FG 3 -3 1-2 -1-3 -2-2 2o ·3·2 -3-2 -4-2 3o o-3 o-4 -3-2 o-4 -1-2 -36 -2 1A - 2 -3 11 I -3 -3 -2 -2 -3 -3 -3 -3 -2-11 -2-10 -20 o -3-2-2 -3-26 -2 iklmnpqrstv wy 12112312221 &gt;- IIIIIIIIIII 2 7 33214423323 _. _. 11111111.11 11 1 3-2!-3-2-2-3-204 •217- 142769.doc 201021828 Table IV: HLA I/II Class Primitives/Super Primitive Table IV (A ) : HLA class I super primitives / primitives

Super element position position 2 (-level anchor point) 3 (-level anchor point) C end (first stage anchor point) A1 TILVMS FWY A2 lAVMATQ YVMATL A3 VSMA7X7 RK A24 YFWIVLMT FIYWLM B7 P VILFMWYA B27 RHK FYLWMIVA B44 ED FWYLIMVA B58 ATS FWYLIVMA B62 QLIVMP YWYMIVLA Primitive A1 TSM Y A1 DIL45 Y A2.1 imVQIAT \LIMAT A3 LMVISATFCGD KYRWE4 All VTMLISAGNCDF KRYH A24 YFWM FLIW A*310 1 MVT^Z/5 RK A*330 1 MVALF/ST RK A*680 1 PNTMSL1 RK B*0702 P IMFWYAIV B*350 1 P LMFWY/B51 P IA\¥WYAM B*530 1 P IMFWY^IF B*540 1 P ATWLMFWY Bold residue is preferred, italic residue is suboptimal: if peptide A peptide having a motif or a super-primary as defined in the above table at each level of anchorage is considered to have a motif. Table IV (B): HLA class II supermotifs

1 6 9 W, F, V, V, .I, L Α, Υ, Ι, L, P, C, S, TA, V, I, L, C, S, T, Μ, Y 142769.doc - 218- 201021828 Table IV (C): HLA Class II Primitives

Elementary level anchor point 1 2 3 4 5 Level 1 anchor point 6 7 8 9 DR4 Better VUYLIVW Μ TI VSTCPALIM MH MH Harmful WR WDE DR1 Better MYLIVWY PAMQ VUATSPLIC M AVM Harmful C CH FD CWD GDE D DR7 Better MYLIVWY Μ WA WMSACTPL M IV Harmful CG GRD NG DR3 Elementary level anchor point 1 2 3 Level 1 anchor point 4 5 Level 1 anchor point 6 Preferred element a LIVMFY D Preferred element b LIVMFAY DNQEST KRH DR super element MYLIVWY VMSTACPLI

Italicized residues indicate suboptimal or "acceptance" residues Table IV (D): HLA Class I supermotif positions: 1 2 3 4 5 6 7 8 C-terminal super-primary A1 First-order error-point one-stage pin point TILVMS FWY A2 First-order error point First-level point LIVlVL4rC? LIVMAT A3 Preferred first-order point VSMA7Z/YFW (4/5) YFW (3/5) YFW (4/5) P (4/5) First-order error RK Harmful DE DE (3 /5); P(5/5) (4/5) A24 Level 1 point level gold m YFWIVLMT FIYWLM B7 Better FWY Level 1 point FWY FWY Level 1 point (5/5) LIVM P (4/5) (3 /5) VILFMWYA Harmful (3/5) DE DE G QN DE (3/5); P(5/5); G(4/5); A(3/5); QN(3/5) (3 /5) (4/5) (4/5) (4/5) ❹ 142769.doc •219- 201021828

B27 First-level wrong point first-level sales point RHK ¥YLWMIVA B44 First-level anchor point-level pin point ED FWYLIMVA B58 - level anchor point level one point error ATS VW^LIVMA B62 First-level point point level pin point QLIVMP FWIMIVLA Italic residue indicates sub-optimal or "resistant Residue Table IV (E): HLA Class I Primitives

Position 1 2 3 4 5 6 7 8 9 C-end or C-end A1 Better GFYW Level 1 DEA YFW P DEQN YFW - Grade 9-mer 锵ft «ft Harmful DE STM RHKLIVMP AGAY Spring A1 Better GRHK ASTCLIVM GSTC ASTC LIVM DE Level 1 9-mer DEAS 锵ft Y Harmful A RHKDEPYFW DE PQN RHK PG GP A1 Better YFW Level 1 DEAQN A YFWQN PASTC GDE P Level 10 -mer 锵ft STM Y Harmful GP RHKGLIVM DE RHK QNA RHKYFW RHK A A1 Preferred YFW STCL1VM - Grade A YFW PG G YFW First-class 10-mer DEAS Y Harmful RHK RHKDEPYFW PG PRHK QN A2.1 Better YFW First-class YFW STC YFW AP - Grade 9-mer 锵 ft 有害 Defective DEP UAIVQAT DERKH RKH DERKH \LJMAT A2.1 AYFW Level 1 LVIM GG FYWLVIML - Grade 10 - mer 锵 ft 锵 ft Harmful DEP IMIVQAT DE RKHA P RKH DERKH RKH WLIMAT A3 Preferred RHK - Grade YFW PRHKYFW A YFW P Level 1 鶬 ft LMVISATFCGD 锵 Post KYKHFA 有害 Harmful DEP DE All Better A First grade YFW YFW A YFW YFW P - grade Α贴«Α VTLMISAGNCZ)F KRYH Harmful DEP AG A24 Preferred YFWRHK First Class STC YFW YFW - Grade 9-mer «ft 锵ft YFWA/· FLIW Harmful DEG DE G QNP DHRHK G AQN A24 Preferred Level P YFWP P - Level 10 -mer Υ典Υ¥ΨΜ FLIW Harmful GDE QN RHK DE A QN DEA A3101 Preferred RHK First Class YFW P YFW YFW AP First Class Sticky UVTALJS RA: Harmful DEP DE ADE DE DE DE A3301 Better - Grade YFW AYFW - Level Error Ft MVALVIST RK Harmful GP DE -220- 142769.doc 201021828

Location 1 2 3 4 5 6 7 8 9 C-end or C-end A6801 Better YFWSTC First-class YFWLIVM YFW P One UL Harmful GP «ft AVT dish / DEG RHK A RK point RK B0702 Better RHKFWY - Grade RHK RHK RHK RHK PA - level harmful DEQNP error ft P DEP DE DE GDE QN DE At code IMYWYAIV B3501 preferred FWYLIVM first level FWY FWY - level harmful AGP PGG for A LMFWY like B51 better LIVMFWY - level FWY STC FWY G FWY _ level harmful AGPDER HKSTC wrong Point P DE G DEQN GDE UMVWYAM B5301 Preferred LIVMFWY - Grade FWY STC FWY LIVMFWY FWY - Grade harmful AGPQN Curved PG RHKQN DE 钃A IMFWY^IK ^5401 Better harmful FWY GPQNDE - Class 锵 It P FWYLIVM GDESTC LIVM RHKDE DE ALIVM QNDGE FWYAP DE - Grade 锵 ft AnVLMFfVY Table IV (F) __ Summary of HLA supertypes Overall phenotype of HLA supertypes of different populations Frequency-specific phenotype Frequency supertype location 2 C-terminal Caucasian North American black 曰 (Caucasian) (NABlack) Chinese Chinese Hispanic Average (Hispanic) B7 P AILMVFWY 43.2 55.1 57.1 43.0 49.3 49.5 A3 AILMVST RK 37.5 42.1 45.8 52.7 43.1 44.2 A2 AILMVT AILMVT 45.8 39.0 42.4 45.9 43.0 42.2 A24 YF (WIVLMT) FI (YWLM) 23.9 38.9 58.6 40.1 38.3 40.0 Q B44 E(D) FWYLIMVA 43.0 21.2 42.9 39.1 39.0 37.0 A1 TI(LVMS) FWY 47.1 16.1 21.8 14.7 26.3 25.2 B27 RHK FYL (WMI) 28.4 26.1 13.3 13.9 35.3 23.4 B62 QL(IVMP) FWY(MIV) 12.6 4.8 36.5 25.4 11.1 18.1 B58 ATS FWY(LIV) 10.0 25.1 1.6 9.0 5.9 10.3 221- 142769.doc 201021828 Table IV (G): I. Population coverage by different HLA supertype combinations HLA supertype A2, A3 &amp; B7 A2, A3, B7, A24, B44 and A1 A2, A3, B7, A24, B44, A1, ίγ ir 『r B27, B62 and B58 Table V: Frequently occurring bases&lt;

16% PH domain, the composition of the closed pleckstrin homology

EGF 34% EGF-like domain

Rvt 49% reverse transcriptase (RNA-dependent DNA polymerase) 30-40 amino acid lengths are present in the extracellular money of the membrane pallidus or secreted by oxygen 4 匕 reduction _ql (Oxidoredql) 25% Ank The repeat sequence NADH·Pan Roll/Quality 32% Awakening (Complex I), different chain cytoplasmic proteins, binds the overall skeleton to the _____ binding membrane. Proton shift across the membrane 142769.doc -222- 201021828 EF Hand (Efhand) Rvp 24% 79% EF hand i transcriptional amine thiol protein 醢 ' ¥ original protein - the binding domain composed of the residue loop, 12 residue loops are flanked by 12-residue α-helical domains _____ aspartame or aspartic acid protease, with catalytic aspartame 醯 a residue as the center of collagen 42% strand repeat Sequence (2 copies) An extracellular structural protein involved in the formation of connective tissue. This sequence consists of G-X-Y and the polypeptide chain forms a triple helix.

The Fn3 20% type III fibronectin domain is located in the extracellular ligand binding region of the receptor and is about 200 amino acid residues in length, of which two pairs of cysteine are implicated by a diterpene bond of 7tm 19% 7 span. Membrane receptor (rhodopsin family) Seven hydrophobic transmembrane regions, where the N-terminus is located extracellularly and the C-terminus is located within the cytoplasm. Signal via G protein _ ❹ Table VI: Determination of affinity based on Ha8-4c4.1

Table VI(A): Ha8-4c4.1 FACS MFI for 3T3-58P1D12 cells

Ha8-4c4.1 MAb Concentration (nM) 58P1D12-3T3 (MFI) 80 1198 40 1167 20 _ 1185 10 1052 5.0 828 2.5 558 1.3 340 0.625 201 0.313 110 0.156 58 0.0781 30 0.0391 18 0.0195 8 0.0098 3 0.0049 0

Table VI(B): Affinity value calculated by Graphpad Prisim software Ha8-4c4.1 saturation curve

-223 - 142769.doc 201021828 BMAX (MFI, 1296 KD(nM) 3.0 [Simplified Schematic] Circle 1A Figure 1A shows the cDNA and amino acid of 58P1D12 variant 1 (also known as "58P1D12 v. 1") Sequence. The initial methionine is underlined. The open reading frame extends from nucleic acid 380 to nucleic acid 1201, including the stop codon; circle 1B is shown in Figure 1B: ^58P1D12 variant 2 (also known as "58P1D12 ν·2" cDNA and amino acid sequence. The codon of the starting thiol amide is underlined. The open reading frame extends from nucleic acid 388 to nucleic acid 1086, including the stop codon; circle 1C Figure 1C shows 58P1D12 variant 3 (also The cDNA and amino acid sequence referred to as "58P1D12 v.3"). The codon for the initiating methionine is underlined. The open reading frame extends from nucleic acid 206 to nucleic acid 904, including the stop codon; Figure 1D Figure 1D The cDNA and amino acid sequence of 58P1D12 variant 4 (also known as "58P1D12 ν·4") is shown. The codon of the starting thiol amide is underlined. The open reading frame extends from nucleic acid 206 to nucleic acid 916, including Stop codon. Figure 1E shows the 58P1D12 variant 5 in Figure 1E (also The cDNA and amino acid sequence of r58pmi2 ν·5"). The codon of the starting thiol amide is underlined. The open reading frame extends from nucleic acid 106 to nucleic acid 816, including the stop codon. Circle IF 58P1D12 v_6 to v .15 (SNP variant of 58P1D12 vl). 142769.doc -224- 201021828 Variant 58P1D12 ν·6 to v.15 are variants with a single nuclear phytic acid difference from 58P1D12 vl. Although these SNP variants are displayed separately However, it may also be present in any combination and in the form of any of the transcript variants listed above in Figure 1 A-1E. Circle 2 The nucleic acid and amino acid sequence of the 58P1D12 antibody. Circle 2A Ha8_4c4· 1 VH cDNA and amine group Acid sequence. The leader sequence is double underlined, and one part of the heavy chain constant region is underlined. Circle 2B Ha8-4c4.1 VL pure 2-A7 cDNA and amino acid sequence. The Yu guide sequence is double underlined, and The light chain constant region is underlined. Circle 2C Ha8-4c4.1 VL pure 1-B3 cDNA and amino acid sequence. The leader sequence is double underlined and the light chain constant region is underlined. Circle 3 58P1D12 antibody (" Amino acid sequence of MAb") Circle 3A Ha8-4 C4.1 Amino acid sequence of VH. The leader sequence is double underlined and one of the heavy chain constant regions is underlined. Loop 3B Ha8-4c4.1 VL pure 2-A7 amino acid sequence. The leader sequence is double underlined and the light chain constant region is underlined. ❹ Figure 3C Ha8-4c4.1 VL pure 1-B3 amino acid sequence. The leader sequence is double underlined and the light chain constant region is underlined. Alignment of the heavy chain variable region of the loop 4 58P1D12 antibody to the human Ig germline.圔 4A Ha8-4c4.1 VH (SEQ ID NO..17) is aligned with the human Ig germline. Circle 4B Ha8-4c4.1 VL pure line 2-A7 (SEQ ID NO: 18) is aligned with the human Ig germ line. Circumference of 4C Ha8-4c4.1 VL pure line 1-B3 (SEQ ID NO: 19) and human Ig 142769.doc -225· 201021828. Loop 5 MAb Ha8-4c4.1 inhibits migration of MDCK cells expressing 58P1D12. Canine MDCK cells were transduced with retrovirus (empty vector [Neo] or 58P1D12). 4x10 of MDCK/58P1D12 cells in 0.1% FBS plus 25 pg/mL control MAb or MAb Ha8-4c4.1 were applied to the chamber above the Boyden Transwell device and allowed to pass into the lower chamber of the cells. % FBS migrated for 16 hours to assess mobility. The cells captured on the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. An empty vector was used to express the cells for use as a negative control. ® quantify the amount of cellular fluorescence (migration) via the MetaMorph imaging software. MAb Ha8-4c4.1 inhibited cell migration by approximately 45%, while the negative control MAb did not inhibit cell migration (*ρ&lt;〇.〇〇〇1). Loop 6 MAb Ha8-4c4.1 inhibits the invasiveness of the OVCAR-5 cells of 58P1D12. Human ovarian cancer cell line OVCAR-5 was transduced with retrovirus (empty vector [Neo] or 58P1D12). The Boyden Transwell chamber was coated with a layer of Matrigel® for cell invasion. MAb Ha8-4c4.1 or an isotype matched control MAb (25 pg/mL) was added to 4 χ 104 OVCAR-5/58P1D12 cells in 0.1% FBS in the stomach of the upper chamber of the Matrigel®-coated device. These cells were allowed to invade for 10 hours with 10% FBS loaded in the lower chamber. Cells bound to the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. Compared to the control MAb, MAb Ha8-4c4.1 significantly inhibited cell invasion by 75% (*ρ&lt;0·0001). Circle 7 compares the active functional activity of 58P1D12 MAb. Testing human cancer with tumor cell migration and tumor cell invasion assay 58P1D12 MAb 142769.doc -226- 201021828

Ha8-4c4.1 (γΐκ), Ha8-6.1 (γ2κ) and Ha8-7.1 (γΐκ). Tumor cell migration was assessed by the Boyden Trangwell chamber migration assay using MDCK/58P1D12 cells. 4 χ 104 of MDCK/58P1D12 cells in 0.1% FBS plus 25 pg/mL control MAb or 58P1D12 MAb were applied to the chamber above the Boyden Transwell device and allowed to migrate 10% of the FBS in the lower chamber. Hours to evaluate the migration. Cells captured on the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. The amount of cellular fluorescence (migration) was quantified by MetaMorph imaging software. Results Table ® shows that Ha8-4c4.1 and Ha8-7.1 MAb inhibit cell migration, while Ha8-6.1 MAb does not inhibit migration. Tumor cell invasion was assessed using a Boyden Transwell chamber coated with a layer of Matrigel® for cell invasion. Briefly, 58P1D12 MAb or isotype matched control MAb (25 pg/mL) was added to 4x104 OVCAR-5/58P1D12 cells in 1·1% FBS in a chamber coated with Matrigel®. These cells were allowed to invade for 10 hours at 10% FBS loaded in the lower chamber. Cells bound to the bottom filter were labeled with calcein AM dye for 30 minutes and photographed. The results showed that 'Ha8-4c4.1 and Ha8-6.1 MAb inhibited tumor cell invasion, while Ha8-7_1 MAb did not inhibit invasion. Circle 8 MAb Ha8-4c4.1 inhibits the formation of the 58P1D12 induced HUVEC tube. Recombinant 58P1D12 ECD (3 pg/mL) was added to HUVEC (5xl〇4/well) in 0.1% FBS with 30 pg/mL isotype matched control MAb or MAb Ha8-4c4.1. The cells were then applied to Matrigel® and allowed to form tubes for 16 hours. Calculate the number of tubes. Control MAb did not affect 142769.doc • 227- 201021828 58P1D12 ECD induced HUVEC tube formation, while MAb Ha8-4c4.1 inhibited tube formation by 50% (*ρ = 0·005). Figure 9 58P1D12 MAb: Comparison of the formation of live HUVEC tubes. Add recombinant 58P1D12 ECD (3 pg/mL) to 30 gg/mL of 58P1D12 MAb Ha8-4c4.1, Ha8_6.1 or Ha8-7.1 to 1111¥£ in 0.1% FBS (:(5&gt;&lt; 104 cells/well. Then the cells were applied to ^^&amp;11^61@ and allowed to form tubes for 16 hours. The number of tubes was calculated. The results showed that all three 58PlD12MAb inhibited tube formation, expressed as (+) Loop 10 for the anti-sputum of 58P1D12 mediates the sand in the 3T3-58P1D12 cells © Bo Ning (83卩01*丨11)-dependent killing effect on the 1st day will be 3-butyl 3-58?1〇12 cells (1000 Cells/wells were seeded in 96-well plates. The next day, a 2-fold concentration of the designated primary antibody was combined with a 2-fold excess of anti-human (Hum_) conjugated to Sabinin toxin (Advanced Targeting Systems, San Diego, CA).

An equal volume of medium of Zap) or anti-goat (Gt Ig Sap) polyclonal antibody was added to each well. The cells were incubated for 4 days at 37 degrees Celsius. At the end of the incubation period, Alamar Blue (Biosource) was added to each well and incubation continued for an additional 4 hours. After excitation at 530 rnn, the fluorescence emission of three heavy G complex samples at 590 nm was determined. The results showed that Ha8-4c4.1 mediates saporin-dependent cytotoxicity in 3T3-58P1D12 cells, whereas the control non-specific human IgGl (H3-1.4.1.2) did not work. These results indicate that drugs or cytotoxic proteins can be selectively delivered to cells expressing 3T3-58P1D12 and other 58P1D12 expressing cells using an appropriate anti-58P1D12 MAb. Study on the efficacy of circle 11 Ha8-4c4.1 in 3T3-58P1D12逋痏. 142769.doc -228- 201021828 3 3-5 8卩1〇12 cells (5,0&gt;&lt;106 cells) were embedded in §61 and the male S CID was implanted at the 0th 小In the right abdomen of the mouse. On the same day, mice were randomized (n=10 per group) and treatment was started intraperitoneally using 500 mg Ha8_4c4.1 or isotype control MAb (twice a week for a total of 8 doses). Tumor growth was monitored using a caliper gauge every 3 to 4 days. As a result, it was confirmed that Hag_4c4.1 inhibited the growth of 3T3-58P1D12 tumor xenografts grown in SCID mice by about 78% as compared with the control antibody alone. The difference in tumor volume between control ® tumors and Ha8_4c4.i tumors was statistically significant (p&lt;0.0001) when analyzed using the Mann-Whitney U test. Figure 12: Effect of Ha8-4e4.1 on established 3T3_58P1D12 tumors grown in mouse tibia. 3T3-58P1D12 cells (5.〇xl〇4 cells) were embedded in Matrigel and operated on the third day. Implanted into the right tibia of male SCID mice. The tumor was established for 7 days, at which time the mice were randomly divided into groups (n=10 per group). Treatment was initiated intraperitoneally using an I.5 mg loading dose of Ha8-4c4.1 or an isotype control 眷 MAb followed by 750 mg of each corresponding MAb (twice per week, total sputum). Tumor growth was monitored using a caliper gauge every 3 to 4 days. The results demonstrated that '24th day' Ha8-4c4.1 inhibited the growth inhibition of established 3T3_58P1di2 tumor xenografts grown in mouse tibia compared to treatment with control antibody (&lt;01.01, Use; Mann-Whitney U test). Circle 13 Ha8-4c4.1 on the efficacy of LAPC-AD prostate • • Enzymatic digestion of LAPC9-AD tumor stock, counting and on the third day will be one hundred 142769.doc • 229- 201021828 500,000 live The cells were implanted subcutaneously into the right tibia of male SCID* mice. On the same day, mice were randomized (n=1 each) and treated with 5 ton of Ha84c4” or isotype control human IgGi in an intraperitoneal manner. Animals were treated twice a week for a total of 10 doses until the end of the study on the 32nd day. The animals were sacrificed and the right tibia and left tibia were weighed via an electronic balance. Tumor weight was determined by subtracting the weight of the tumor-free right tibia from the weight of the contralateral tibia without tumor, and plotting the tumor weight. As a result, it was confirmed that Ha8_4c4.丄 inhibited the growth of LAPC9-AD prostate cancer xenografts grown in the mouse tibia by 60% compared with the control antibody treatment. The difference between the control tumor weight and the Ha8-4c4.1 tumor weight was statistically significant (p=0.0057) when using Student's t-test analysis. Circle 14 Ha8-4c4.1 is effective for ovarian tumors growing in the tibia of mice. Tumor cells (2·〇χ106 cells) expressing Ovcar5-58PlD12 were implanted into the right tibia of female SCID mice. After the sputum, the mice were randomized (n = 1 每 each) and the treatment was initiated intraperitoneally (i.P.) using 5 〇〇 Ha8-4c4.l or isotype control human IgGl. Animals were treated twice a week for a total of 12 doses 'up to 42nd. At the end of the study (p. 42), the animals were sacrificed and the right tibia and left tibia were weighed via an electronic balance. The tumor weight is the measured value obtained after subtracting the weight of the contralateral tibia without the tumor, and the tumor weight is plotted as a graph. The results demonstrated that Ha8-4c4.1 was effective against the 0vcar5-58PlD12 tumor in a single agent form compared to the control antibody treatment, resulting in a 56% inhibition of growth (p=〇.〇〇〇2, using the Mann-Whitney U test). 142769.doc -230- 201021828 The effect of circle 15 Ha8-4c4.1 on the survival of SCID mice with intraperitoneal established vCAR-5-58P1D12 tumors. In the third place, 〇vcar5_58piD12 tumor cells (2·〇χ106 cells) were injected into the peritoneum of female sciD mice. When the tumor was fully established, the mice were randomized into groups (n=15 per group) and treatment was started intraperitoneally using 500 Ha8-4c4.1 or isotype control human IgG1. Animals are treated with antibodies twice a week as long as they survive. The health and survival of the mice were monitored and recorded every few days. The results demonstrated that mice with well established ovarian tumors treated with Ha8-4c4.1 survived a median of 69 days and mice treated with control MAb survived a median of 37 days. The median increase in the number of days of survival in mice treated with Ha8_4c41 was statistically significant (p=0 〇〇66, using L〇gran]&lt;^ test). The effect of the combination of circle 16 1138-4〇4.1舆 card ([31*1)〇口丨3^11). The ability of Ha8_φ 4c4.1 as a monotherapy and in combination with the chemotherapeutic carboplatin was evaluated in an established androgen-independent prostate tumor xenograft (lapc: 9_ai). The stock solution of WLAPC9-AI tumor was enzymatically digested, and 1.5 x 106 cells were surgically implanted into the right tibia of male SCID mice on day 0. The tumors were allowed to establish for 7 days, at which time the animals were randomly divided into four different groups (n=l each for each group) as shown in the graph. Starting on day 7, a loading dose (2 mg) of Ha8-4c4.1 or an isotype control human IgG1 was administered intraperitoneally, followed by a maintenance dose (1.0 mg) of the corresponding MAb, twice weekly for a total of 7 doses. On the 7th, Uth, 15th, 19th, 22nd and 26th, the card (4. mg/kg) was administered intravenously (i.v.) to the mice. On page 33, all mice were sacrificed and the tumors were excised and weighed using an electronic balance. 142769.doc -231 - 201021828 The results demonstrate that Ha8-4c4.1 is very effective in a single agent form and inhibits tumor growth by 76% compared to control antibody treatment (p=0.0077). Cardatic monotherapy also inhibited tumor growth, resulting in inhibition of tumor growth by 87% (p=0.0001). Combination of Ha8_4c4.1 and carboplatin increased, strong inhibition and tumor growth was inhibited by 97% compared with control antibody alone. (p&lt;0.0001). Statistically significant differences were also demonstrated when comparing the tumor weight of the Ha8-4c4.1 plus carboplatin treatment group to the tumor weight of the control MAb plus carboplatin treatment group (p = 〇. 〇 243). Statistical analysis was performed initially using the Kruskal-Wallis test to determine significance in each group. Subsequently, a student t test or a Mann-Whitney U test was applied to each pair. 142769.doc -232- 201021828 Sequence Listing &lt;110>American Azei Company &lt;120&gt; antibody and related molecule that binds to 58P1D12 protein &lt;130&gt; 511582002082 &lt;140> 098128076 &lt;141&gt; 2009-08- 20 &lt;150&gt; 61/207,862 &lt;151> 2008-08-20 &lt;150&gt; 61/153,225 &lt;151> 2009-02-17 &lt;160> 19 &lt;170&gt; FastSEQ for Windows Version 4. 0

&lt;210&gt; 1 &lt;211&gt; 2550 &lt;212> DNA &lt;2i3&gt; Homo sapiens &lt;220&gt;

&Lt; 221 &gt; CDS &lt; 222> (380) ... (1201) &lt; 400> 1 atccaggacc agggcgcacc ggctcagcct ctcacttgtc agaggccggg gaagagaagc 60 aaagcgcaac ggtgtggtcc aagccggggc ttctgcttcg cctctaggac atacacggga 120 ccccctaact tcagtccccc aaacgcgcac cctcgaagtc ttgaactcca gccccgcaca 180 tccacgcgcg gcacaggcgc ggcaggcggc aggtcccggc cgaaggcgat gcgcgcaggg 240 ggtc-gggcag ctgggctcgg gcggcgggag tagggcccgg cagggaggca gggaggctgc 300 agagtcagag tcgcgggctg cgccctgggc agaggccgcc ct-cgctccac gcaacacctg 360 ctgctgccac cgcgccgcg atg age ege gtg gtc teg ctg ctg ctg ggc gee 412 Met Ser Arg Val Val Ser Leu Leu Leu Gly Ala 1 5 10 geg ctg etc Tgc ggc cac gga gee ttc tgc ege ege gtg gtc age ggc 460

Ala Leu Leu Cys Gly His Gly Ala Phe Cvs Arg Arg Val Val Ser Gly 15 20 25 c-aa aa.g gtg tgt ttt get gac ttc aag cat ccc tgc tac aaa atg gee 508

Gin Lys Val Cys Phe Ala Asp Phe Lys His Pro Cys Tvr Lys Met Ala 30 35 40

Tac ttc cat gaa ctg tee age ega gtg age ttt cag gag gca ege ctg 556

Tyr Phe His Glu Leu Ser Ser Arg Val Ser Phe Gin Glu Ala Arg Leu 45 50 55 get tgt gag agt gag gga gga gtc etc etc age ett gag aat. gaa gca 604

Ala Cvs Glu Ser Glu Gly Gly Val Leu Leu Ser Leu Glu Asn Glu Ala 60 65 70 75 gaa cag aag tta ata gag age atg ttg caa aac ctg aca aaa ccc ggg 652

Glu Gin Lys Leu lie Glu Ser Met Leu Gin Asu Leu Tlir Lys Pro Gly 80 85 90 aca ggg att tet gat ggt gat ttc tgg ata ggg ett tgg agg aat gga 700

Thr Gly lie Ser Asp Gly Asp Phe Trp He Glv Leu Trp Arg Asn Gly 95 100 ^ 105 gat ggg caa aca tet ggt gee tgc cca gat etc tac cag tgg tet gat 748

Asp Gly Gin Thr Ser Gly Ala Cvs Pro Asp Leu Tyr Gin Trp Ser Asp 110 115 120 142769 - Sequence Listing.doc 201021828 gga age aat tee cag tac ega aac tgg tac aca gat gaa cct tee *tgc 796

Gly Ser Asn Ser Gin Tyr Arg Asn Trp Tyr Thr Asp Glu Pro Ser Cys 125 130 135 gga agt gaa aag tgt gtt gtg atg tat cac caa cca act gee aat c:ct 844

Gly Ser Glu Lys Cys Val Val Met Tyr His Gin Pro Thr Ala Asn Pro 140 145 150 155 ggc ett ggg ggt ecc tac ett tac cag tgg aat gat gac agg tgt aac 892

Gly Leu Gly Gly Pro Tyr Leu Tyr Gin Trp Asn Asp Asp Arg Cys Asn 160 165 170 atg aag cac aat tat att tgc aag tat gaa cca gag att aat cca aca 940

Met Lys His As Tyr lie Cys Lys Tyr Glu Pro Glu He As Pro Pro 175 180 185 gee cct gta gaa aag cct tat ett aca aat caa cca gga gac acc cat 988

Ala Pro Val Glu Lys Pro Tyr Leu Thr Asn Gin Pro Gly Asp Thr His 190 195 200 cag aat gtg gtt gtt act gaa gca ggt ata att ccc aat eta att tat 1036

Gin Asn Val Val Val Thr Glu Ala Gly lie lie Pro Asn Leu lie Tyr 205 210 215 gtt gtt ata cca aca ata ccc ctg etc tta ctg ata ctg gtt get t:tt 1084

Val Val lie Pro Thr lie Pro Leu Leu Leu Leu lie Leu Val Ala Phe 220 225 230 2 35 gga acc tgt tgt ttc cag atg ctg cat aaa agt aaa gga aga aca aaa 1132

Gly Thr Cys Cys Phe Gin Met Leu His Lys Ser Lys Gly Arg Thr Lys 240 245 250 act agt cca aac cag tet aca ctg tgg att tea aag agt acc aga a.aa 1180

Thr Ser Pro Asn Gin Ser Thr Leu Trp lie Ser Lys Ser Thr Arg Lys 255 260 265 gaa agt ggc atg gaa gta taa taactcattg acttggttcc agaattttgt 1231

Glu Ser Gly Met Glu Val * 270 aattctggat ctgtataagg aatggcatca gaacaatagc ttggaatggc ttgaaatcac 1291 aaaggatctg caagatgaac tgtaagctcc cccttgaggc aaatattaaa gtaattttta 1351 tatgtetatt atttcattta aagaatatgc tgtgctaata atggagtgag acatgcttat 1411 tttgctaaag gatgcaccca aacttcaaac ttcaagcaaa tgaaatggac aatgeagata 1471 aagttgttat caacacgtcg ggagtatgtg tgttagaagc aattcctttt atttctttca 1531 cctttcataa gttgttatct agtcaatgta atgtatattg tattgaaatt tacagtgtgc 1591 aaaagtattt tacctttgca taagtgtttg ataaaaatga actgttctaa tattta.tttt 1651 tatggeatet catttttcaa tacatgctct tttgattaaa gaaacttatt actgtt: gtca 1711 actgaattca cacacacaca aatatagtac catagaaaaa gtttgttttc tcgaaataat 1771 teatetttea gcttctctgc ttttggtcaa tgtctaggaa atctcttcag aaataagaag 1831 ctatttcatt aagtgtgata taaacctcct caaacatttt aettagagge aagga * ttgtc 1891 taatttcaat tgtgcaagac atgtgcctta taattatttt tagcttaaaa ttaaaoagat 1951 tttgtaataa tgtaactttg Tatataggg cataaacact aatgeagtea atttgaacaa 2011 aagaagtgac atacacaata taaatcatat gtcttcacac gttgcctata taatgageiag 2071 cagctctctg agggttctga aatcaatgtg gtccctctct tgcccactaa acaaagatgg 2131 ttgttcgggg tttgggattg acactggagg cagatagttg caaagttagt ctaaggtttc 2191 cctagctgta tttagcctct gactatatta gtatacaaag aggtcatgtg gttgagacca 2251 ggtgaatagt cactatcagt gtggagacaa gcacagcaca cagacatttt aggaaggaaa 2311 ggaactacga aatcgtgtga aaatgggttg gaacccatca gtgatcgcat attcat: tgat 2371 gagggtttgc ttgagataga aaatggtggc tcctttctgt cttatctcct agtttcttca 2431 atgettaege cttgttcttc tcaagagaaa Gttgtaactc tctggtcttc atatg*tccct 2491 gtgctccttt taaccaaata aagagttctt gtttctgaag aaaaaaaaaa aaaaaaaaa 2550 &lt;210〉 2 &lt;211> 273 &lt;212&gt; PRT &lt;213> Homo sapiens &lt;400〉 2

Met Ser Arg Val Val Ser Leu Leu Leu Gly Ala Ala Leu Leu Cys Gly 15 10 15

His Gly Ala Phe Cys Arg Arg Val Val Ser Gly Gin Lys Val Cys Phe 20 25 30

Ala Asp Phe Lys His Pro Cys Tyr Lys Met Ala Tyr Phe His Glu Leu • 2· 142769· Sequence Listing.doc 201021828 35 40 45

Ser Ser Arg Val Ser Phe Gin Glu Ala Arg Leu Ala Cys Glu Ser Glu 50 55 60

Gly Gly Val Leu Leu Ser Leu Glu Asn Glu Ala Glu Gin Lys Leu lie 65 70 75 80

Glu Ser Met Leu Gin Asn Leu Thr Lys Pro Gly Thr Gly lie Ser Asp 85 90 95

Gly Asp Phe Trp lie Gly Leu Trp Arg Asn Gly Asp Gly Gin Thr Ser 100 105 110

Gly Ala Cys Pro Asp Leu Tyr Gin Trp Ser Asp Gly Ser Asn Ser Gin 115 120 125

Tyr Arg Asn Trp Tyr Thr Asp Glu Pro Ser Cys Gly Ser Glu Lys Cys 130 135 140

Val Val Met Tyr His Gin Pro Thr Ala Asn Pro Gly Leu Gly Gly Pro 145 150 155 160

Tyr Leu Tyr Gin Trp Asn Asp Asp Arg Cys Asn Met Lys His Asn Tyr 165 170 175 lie Cys Lys Tyr Glu Pro Glu lie Asn Pro Thr Ala Pro Val Glu Lys 180 185 190

Pro Tyr Leu Thr Asn Gin Pro Gly Asp Thr His Gin Asn Val Val Val 195 200 205

Thr Glu Ala Gly lie lie Pro Asn Leu lie Tyr Val Val lie Pro Thr 210 215 220 lie Pro Leu Leu Leu Leu lie Leu Val Ala Phe Gly Thr Cys Cys Phe 225 230 235 240

Gin Met Leu His Lys Ser Lys Gly Arg Thr Lys Thr Ser Pro Asn Gin 245 250 255

Ser Thr Leu Trp lie Ser Lys Ser Thr Arg Lys Glu Ser Gly Met Glu 260 265 270

Val &lt;210> 3 &lt;211> 2418 &lt;212> DNA &lt;213> Homo sapiens &lt;220〉

&lt;221> CDS &lt;222>(388)... (1086) &lt;400〉 3

ctgtggtgtt tttcccccgc tcctctggct gccttcctga tggatctctg tggtcccagg 60 caggaatggc ctgcttgggg acccagcgag ctcccaaggc ctttcctgct gcttcctcta 120 tccctgtgtt ttgcttggct ctctaaattg actcagctcc aggacatcag gaccccaggt 180 tctctggtct tgggactctg agacttgcac caggaatcct gcccaggctc tcaggccttt 240 ggactcagac tgagctactt cactggcttt cctggttctc cagcttgaag atggcagatc 300 gtgggacttc tcagcctcca taattgagtg agccaattcc ctggccaaaa ggtgtgtttt 360 gctgacttca agcatccctg ctacaaa atg gcc tac ttc cat Gaa ctg tcc age 414

Met Ala Tyr Phe His Glu Leu Ser Ser ega gtg age ttt cag gag gca ege ctg get tgt gag agt gag gga gga 462

Arg Val Ser Phe Gin Glu Ala Arg Leu Ala Cys Glu Ser Glu Gly Gly 10 15 20 25 gtc etc etc age ett gag aat gaa gca gaa cag aag tta ata gag age 510

Val Leu Leu Ser Leu Glu Asn Glu Ala Glu Gin Lys Leu lie Glu Ser 30 35 40 atg ttg caa aac ctg aca aaa ccc ggg aca ggg att tet gat ggt gat 558

Met Leu Gin Asn Leu Thr Lys Pro Gly Thr Gly lie Ser Asp Gly Asp 45 50 55 ttc tgg ata ggg ett tgg agg aat gga gat ggg caa aca tet ggt gcc 606

Phe Trp lie Gly Leu Trp Arg Asn Gly Asp Gly Gin Thr Ser Gly Ala 60 65 70 tgc cca gat etc tac cag tgg tet gat gga age aat tcc cag tac ega 654

Cys Pro Asp Leu Tyr Gin Trp Ser Asp Gly Ser Asn Ser Gin Tyr Arg 75 80 85 aac tgg tac aca gat gaa cct tcc tgc gga agt gaa aag tgt gtt gtg 702

Asn Trp Tyr Thr Asp Glu Pro Ser Cys Gly Ser Glu Lys Cys Val Val 142769 · Sequence Listing.doc 201021828 90 95 ΙΟΟ 105 atg tat cac caa cca act gcc aat cct ggc ct匕 ggg ggt ccc tac ctt 750

Met Tyr His Gin Pro Thr Ala Asn Pro Gly Leu. Gly Gly Pro Tyr Leu 110 115 120 tac cag tgg aat gat gac agg tgt aac atg aa^ cac aat tat att tgc 798

Tyr Gin Trp Asn Asp Asp Arg Cys Asn Met Lys His Asn Tyr lie Cys 125 130 135 aag tat gaa cca gag att aat cca aca gcc cct gta gaa aag cct tat 846

Lys Tyr Glu Pro Glu lie Asn Pro Thr Ala Pro Val Glu Lys Pro Tyr 140 145 150 ctt aca aat caa cca gga gac acc cat cag aat gtg gtt gtt act gaa 894

Leu Thr Asn Gin Pro Gly Asp Thr His Gin Asn. Val Val Val Thr Glu 155 160 165 gca ggt ata att ccc aat eta att tat gtt gtt ata cca aca ata ccc 942

Ala Gly lie lie Pro Asn Leu lie Tyr Val VaL lie Pro Thr lie Pro 170 175 180 185 ctg etc tta ctg ata ctg gtt get ttt gga acc tgt tgt ttc cag atg 990

Leu Leu Leu Leu lie Leu Val Ala Phe Gly Thr- Cys Cys Phe Gin Met 190 195 200 ctg cat aaa agt aaa gga aga aca aaa act agt, cca aac cag tet aca 1038

Leu His Lys Ser Lys Gly Arg Thr Lys Thr Ser- Pro Asn Gin Ser Thr 205 210 215 ctg tgg att tea aag agt acc aga aaa gaa agfc ggc atg gaa gta taa 1086

Leu Trp lie Ser Lys Ser Thr Arg Lys Glu Ser * Gly Met Glu Val * 220 225 230 taactcattg acttggttcc agaattttgt aattctggat ctgtataagg eLatggcatca 1146 gaacaatagc ttggaatggc ttgaaatcac aaaggatctg caagatgaac tgtaagctcc 1206 gatgcaccca aacttcaaac 1326 cccttgaggc aaatattaaa gtaattttta tatgtetatt atttcattta aagaatatgc 1266 tgtgctaata atggagtgag acatgcttat tttgctaaag ttcaagcaaa tgaaatggac aatgeagata aagttgttats caacacgtcg ggagtatgtg 1386 tgttagaagc aattcctttt atttctttca cctttcataa, gttgttatct agtcaatgta 1446 atgtatattg tattgaaatt tacagtgtgc aaaagtattt tacctttgca taagtgtttg 1506 ataaaaatga actgttctaa tatttatttt tatggeatet: catttttcaa tacatgctct 1566 tttgattaaa gaaacttatt actgttgtca actgaattcaL cacacacaca aatatagtac 1626 catagaaaaa gtttgttttc tcgaaataat tcatctttcaL gcttctctgc ttttggtcaa 1686 tgtctaggaa atctcttcag aaataagaag ctatttcatt aagtgtgata taaacctcct 1746 caaacatttt Aettagagge aaggattgtc taatttcaat: tgtgcaagac atgtgcctta 1806 taattatttt tagcttaaaa tta£Lacagat tttgtaataa- tgtaactttg ttaatagg tg 1866 cataaacact aatgeagtea atttgaacaa aagaagtga &amp; atacacaata taaatcatat 1926 gtcttcacac gttgcctata taatgagaag cagctctctg agggttctga aatcaatgtg 1986 gtccctctct tgcccactaa acaaagatgg ttgttcgggg:. tttgggattg acactggagg 2046 cagatagttg caaagttagt ctaaggtttc cctagctgta tttagcctct gactatatta 2106 gtatacaaag aggtcatgtg gttgagacca ggtgaatagt: cactatcagt gtggagacaa 2166 gcacagcaca cagacatttt aggaaggaaa ggaactacga, aatcgtgtga aaatgggttg 2226 gaacccatca gtgatcgcat attcattgat gagggtttgc: ttgagataga aaatggtggc 2286 tcctttctgt cttatctcct agtttcttca atgettaege- cttgttcttc tcaagagaaa 2346 gttgtaactc tctggtcttc atatgtccct gtgctccttt taaccaaata aagagttctt 2406 gtttctgaag aa 2418 &lt; 210 &gt; 4 &lt; 211> 232 &lt; 212 &gt; PRT &lt; 213> Homo sapiens &lt; 400 &gt; 4

Met Ala Tyr Phe His Glu Leu Ser Ser Arg VaL Ser Phe Gin Glu Ala 1 5 10 15

Arg Leu Ala Cys Glu Ser Glu Gly Gly Val Lem Leu Ser Leu Glu Asn 20 25 30

Glu Ala Glu Gin Lys Leu lie Glu Ser Met Lett Gin Asn Leu Thr Lys 35 40 45

Pro Gly Thr Gly lie Ser Asp Gly Asp Phe Trp lie Gly Leu Trp Arg 50 55 60

Asn Gly Asp Gly Gin Thr Ser Gly Ala Cys Pro Asp Leu Tyr Gin Trp 65 70 75 80

Ser Asp Gly Ser Asn Ser Gin Tyr Arg Asn Trp Tyr Thr Asp Glu Pro -4- 142769 · Sequence Listing.doc 201021828 85 90 95

Ser Cys Gly Ser Glu Lys Cys Val Val Met Tyr His Gin Pro Thr Ala 100 105 110

Asn Pro Gly Leu Gly Gly Pro Tyr Leu Tyr Gin Trp Asn Asp Asp Arg 115 120 125

Cys Asn Met Lys His As Tyr lie Cys Lys Tyr Glu Pro Glu lie Asn 130 135 140

Pro Thr Ala Pro Val Glu Lys Pro Tyr Leu Thr Asn Gin Pro Gly Asp 145 150 155 160

Thi&quot; His Gin Asn Val Val Val Thr Glu Ala Gly lie lie Pro Asn Leu 165 170 175 lie Tyr Val Val lie Pro Thr lie Pro Leu Leu Leu Leu lie Leu Val 180 185 190

Ala Phe Gly Thr Cys Cys Phe Gin Met Leu His Lys Ser Lys Gly Arg 195 200 205

Thr Lys Thr Ser Pro Asn Gin Ser Thr Leu Trp lie Ser Lys Ser Thr 210 215 220

Arg Lys Glu Ser Gly Met Glu Val 225 230 &lt;210&gt; 5 &lt;211> 2236 &lt;212> DNA &lt;213> Homo sapiens

&Lt; 221> CDS &lt; 222> (206) · .. (904) &lt; 400> 5 gcttgaagat ggcagatcgt gggacttctc agcctccata attgagtgag ccaattccct 60 gaatacaaca agaagatggc catctgtaga ccaggaggtg gtccctcccc agaaactgga 120 tgggccagca cgttgattct gagcttctag cctccagaac tgccaaaagg tgtgttttgc 180 tgacttcaag catccctgct acaaa atg gcc tac Ttc cat gaa ctg tcc age 232

Met Ala Tyr Phe His Glu Leu Ser Ser ega gtg age ttt cag gag gca ege ctg get tgt gag agt gag gga gga 280

Arg Val Ser Phe Gin Glu Ala Arg Leu Ala Cys Glu Ser Glu Gly Gly 10 15 20 25 gtc etc etc age ett gag aat gaa gca gaa cag aag tta ata gag age 328

Val Leu Leu Ser Leu Glu Asn Glu Ala Glu Gin Lys Leu lie Glu Ser 30 35 40 atg ttg caa aac ctg aca aaa ccc ggg aca ggg att tet gat ggt gat 376

Met Leu Gin Asn Leu Thr Lys Pro Gly Thr Gly lie Ser Asp Gly Asp 45 50 55 ttc tgg ata ggg ett tgg agg aat gga gat ggg caa aca tet ggt gcc 424

Phe Trp lie Gly Leu Trp Arg Asn Gly Asp Gly Gin Thr Ser Gly Ala 60 65 70 tgc cca gat etc tac cag tgg tet gat gga age aat tcc cag tac ega 472

Cys Pro Asp Leu Tyr Gin Trp Ser Asp Gly Ser Asn Ser Gin Tyr Arg 75 80 85 aac tgg tac aca gat gaa cct tcc tgc gga agt gaa aag tgt gtt gtg 520

Asn Trp Tyr Thr Asp Glu Pro Ser Cys Gly Ser Glu Lys Cys Val Val 90 95 100 105 atg tat cac caa cca act gcc aat cct ggc ett ggg ggt ccc tac ett 568

Met Tyr His Gin Pro Thr Ala Asn Pro Gly Leu Gly Gly Pro Tyr Leu 110 115 120 tac cag tgg aat gat gac agg tgt aac atg aag cac aat tat att tgc 616

Tyr Gin Trp Asn Asp Asp Arg Cys Asn Met Lys His Asn Tyr lie Cys 125 130 135 aag tat gaa cca gag att aat cca aca gcc cct gta gaa aag cct tat 664

Lys Tyr Glu Pro Glu lie Asn Pro Thr Ala Pro Val Glu Lys Pro Tyr 140 145 150 ett aca aat caa cca gga gac acc cat cag aat gtg gtt gtt act gaa 712 142769 · Sequence Listing.doc 201021828

Leu Thr Asn Gin Pro Gly Asp Thr His Gin Asn Val Val Val Thr Glu 155 160 165 gca ggt ata att ccc aat eta att tat gtt gtt ata cca aca ata ccc 760

Ala Gly lie lie Pro Asn Leu lie Tyr Val Val lie Pro Thr lie Pro 170 175 180 185 ctg etc tta ctg ata ctg gtt get ttt gga acc tgt tgt ttc cag atg 808

Leu Leu Leu Leu lie Leu Val Ala Phe Gly Thr Cys Cys Phe Gin Met 190 195 200 ctg cat aaa agt aaa gga aga aca aaa act agt cca aac cag tet aca 856

Leu His Lys Ser Lys Gly Arg Thr Lys Thr Ser Pro Asn Gin Ser Thr 205 210 215 ctg tgg att tea aag agt acc aga aaa gaa agt ggc atg gaa gta taa 904

Leu Trp He Ser Lys Ser Thr Arg Lys Glu Ser Gly Met Glu Val * 220 225 230 taactcattg acttggttcc agaattttgt aattctggat ctgtataagg aatggcatca 964 gaacaatagc ttggaatggc ttgaaatcac aaaggatctg caagatgaac tgtaagctcc 1024 cccttgaggc aaatattaaa gtaattttta tatgtetatt atttcattta aagaatatgc 1084 tgtgctaata atggagtgag acatgcttat tttgctaaag gatgcaccca aacttcaaac 1144 ttcaagcaaa tgaaatggac aatgeagata aagttgttat caacacgtcg ggagtatgtg 1204 tgttagaagc aattcctttt atttctttca cctttcataa gttgttatct agtcaatgta 1264 atgtatattg tattgaaatt tacagtgtgc aaaagtattt tacctttgca taagtgtttg 1324 ataaaeiatga actgttctaa tatttatttt tatggeatet catttttcaa tacatgctct 1384 tttgattaaa gaaacttatt actgttgtca actgaattca cacacacaca aatatagtac 1444 catagaaaaa gtttgttttc tcgaaataat teatetttea gcttctctgc ttttggtcaa 1504 tgtctaggaa atctcttcag aaataagaag ctatttcatt aagtgtgata taaacctcct 1564 caaacatttt aettagagge aaggattgtc taatttcaat Tgtgcaagac atgtgcctta 1624 taattatttt tagcttaaaa ttaaacagat tttgtaataa tgtaactttg ttaataggtg 1684 cata aacact aatgeagtea atttgaacaa aagaagtgac atacacaata taaatcatat 1744 gtcttcacac gttgcctata taatgagaag cagctctctg agggttctga aatcaatgtg 1804 gtccctctct tgcccactaa acaaagatgg ttgttcgggg tttgggattg acactggagg 1864 cagatagttg caaagttagt ctaaggtttc cctagctgta tttagcctct gactatatta 1924 gtatacaaag aggtcatgtg gttgagacca ggtgaatagt cactatcagt gtggagacaa 1984 gcacagcaca cagacatttt aggaaggaaa ggaactacga aatcgtgtga aaatgggttg 2044 gaacccatca gtgatcgcat attcattgat gagggtttgc ttgagataga aaatggtggc 2104 tcctttctgt Cttatctcct agtttcttca atgettaege cttgttcttc tcaagagaaa 2164 gttgtaactc tctggtcttc atatgtccct gtgctccttt taaccaaata aagagttctt 2224 gtttctgaag aa 2236 &lt;210> 6 &lt;211> 232 &lt;212&gt; PRT &lt;213> Homo sapiens &lt;400&gt; 6

Met Ala Tyr Phe His Glu Leu Ser Ser Arg Val Ser Phe Gin Glu Ala 15 10 15

Arg Leu Ala Cys Glu Ser Glu Gly Gly Val Leu Leu Ser Leu Glu Asn 20 25 30

Glu Ala Glu Gin Lys Leu lie Glu Ser Met Leu Gin Asn Leu Thr Lys 35 40 45

Pro Gly Thr Gly lie Ser Asp Gly Asp Phe Trp lie Gly Leu Trp Arg 50 55 60

Asn Gly Asp Gly Gin Thr Ser Gly Ala Cys Pro Asp Leu Tyr Gin Trp 65 70 75 80

Ser Asp Gly Ser Asn Ser Gin Tyr Arg Asn Trp Tyr Thr Asp Glu Pro 85 90 95

Ser Cys Gly Ser Glu Lys Cys Val Val Met Tyr His Gin Pro Thr Ala 100 105 110

Asn Pro Gly Leu Gly Gly Pro Tyr Leu Tyr Gin Trp Asn Asp Asp Arg 115 120 125

Cys Asn Met Lys His As Tyr He Cys Lys Tyr Glu Pro Glu lie Asn 130 135 140

Pro Thr Ala Pro Val Glu Lys Pro Tyr Leu Thr Asn Gin Pro Gly Asp 145 150 155 160

Thr His Gin Asn Val Val Val Thr Glu Ala Gly lie lie Pro Asn Leu 165 170 175 lie Tyr Val Val lie Pro Thr lie Pro Leu Leu Leu Leu lie Leu Val 180 185 190

Ala Phe Gly Thr Cys Cys Phe Gin Met Leu His Lys Ser Lys Gly Arg 195 200 205 -6 - 142769 - Sequence Listing.doc 201021828

Thr Lys Thr Ser Pro Asn Gin Ser Thr Leu Trp He Ser Lys Ser Thi* 210 215 220

Arg Lys Glu Ser Gly Met Glu Val 225 230 &lt;210&gt; 7 &lt;211> 2133 &lt;212> DNA &lt;213> Homo sapiens &lt;220〉

&Lt; 221> CDS &lt; 222 &gt; (206) ... (916) &lt; 400> 7 gcttgaagat ggcagatcgt gggacttctc agcctccata attgagtgag ccaattccct 60 gaatacaaca agaagatggc catctgtaga ccaggaggtg gtccctcccc agaaactgga 120 tgggccagca cgttgattct gagcttctag cctccagaac tgccaaaagg tgtgttttgc 180 tgacttcaag catccctgct acaaa atg gcc tac Ttc cat gaa ctg tcc age 232

Met Ala Tyr Phe His Glu Leu Ser Ser ega gtg age ttt cag gag gca ege ctg get tgt gag agt gag gga gga 280

Arg Val Ser Phe Gin Glu Ala Arg Leu Ala Cys Glu Ser Glu Gly Gly 10 15 20 25 gtc etc etc age ett gag aat gaa gca gaa cag aag tta ata gag age 328

Val Leu Leu Ser Leu Glu Asn Glu Ala Glu Gin Lys Leu lie Glu Ser 30 35 40 atg ttg caa aac ctg aca aaa ccc ggg aca ggg att tet gat ggt gat 376

Met Leu Gin Asn Leu Thr Lys Pro Gly Thr Gly lie Ser Asp Gly Asp 45 50 55 ttc tgg ata ggg ett tgg agg aat gga gat ggg caa aca tet ggt gcc 424

Phe Trp He Gly Leu Trp Arg Asn Gly Asp Gly Gin Thr Ser Gly Ala 60 65 70 tgc cca gat etc tac cag tgg tet gat gga age aat tec cag tac ega 472

Cys Pro Asp Leu Tyr Gin Trp Ser Asp Gly Ser Asn Ser Gin Tyr Arg 75 80 85 aac tgg tac aca gat gaa cct tec tgc gga agt gaa aag tgt gtt gtg 520

Asn Trp Tyr Thr Asp Glu Pro Ser Cys Gly Ser Glu Lys Cys Val Val 90 95 100 105 atg tat cac caa cca act gcc aat cct ggc ett ggg ggt ccc tac ett 568

Met Tyr His Gin Pro Thr Ala Asn Pro Gly Leu Gly Gly Pro Tyr Leu 110 115 120 tac cag tgg aat gat gac agg tgt aac atg aag cac aat tat att tgc 616

Tyr Gin Trp Asn Asp Asp Arg Cys Asn Met Lys His Asn Tyr lie Cys 125 130 135 aag tat gaa cca gag att aat cca aca gcc cct gta gaa aag cct tat 664

Lys Tyr Glu Pro Glu lie A.sn Pro Thr Ala Pro Val Glu Lys Pro Tyr 140 145 150 ett aca aat caa cca gga gac acc cat cag aat gtg gtt gtt act gaa 712

Leu Thr Asn Gin Pro Gly Asp Thr His Gin Asn Val Val Val Thr Glu 155 160 165 gca gta aag gaa gaa caa aaa eta gtc caa acc agt eta cac tgt gga 760

Ala Val Lys Glu Glu Gin Lys Leu Val Gin Thr Ser Leu His Cys Gly 170 175 180 185 ttt caa aga gta cca gaa aag aaa, gtg gca tgg aag tat aat aac tea 808

Phe Gin Arg Val Pro Glu Lys Lys Val Ala Ti*p Lys Tyr Asn Asn Ser 190 195 200 ttg act tgg ttc cag aat ttt gta att ctg gat ctg tat aag gaa tgg 856

Leu Thr Trp Phe Gin Asn Phe Val lie Leu Asp Leu Tjrr Lys Glu Trp 205 210 215 142769 - Sequence Listing.doc 201021828 cat cag aac aat age ttg gaa tgg ett gaa ate aca aag gat ctg caa 904

His Gin Asn Asn Ser Leu Glu Trp Leu Glu lie Thr Lys Asp Leu Gin 220 225 230 gat gaa ctg taa gctccccctt gaggcaaata ttaaagtaat ttttatatgt 956

Asp Glu Leu * 235 ctattatttc atttaaagaa tatgctgtgc taataatgga gtgagacatg cttattttgc 1016 taaaggatgc acccaaactt caaacttcaa gcaaatgaaa tggacaatgc agataaagtt 1076 gttatcaaca cgtcgggagt atgtgtgtta gaagcaattc cttttatttc tttcaccttt 1136 cataagttgt tatetagtea atgtaatgta tattgtattg aaatttacag tgtgcaaaag 1196 tattttacct ttgcataagt gtttgataaa aatgaactgt tetaatattt atttttatgg 1256 catctcattt ttcaatacat gctcttttga ttaaagaaac ttattactgt tgtcaactga 1316 attcacacac acacaaatat agtaccatag aaaaagtttg ttttctcgaa ataattcatc 1376 tttcagcttc tctgcttttg gtcaatgtct aggaaatctc ttcagaaata agaagetatt 1436 tcattaagtg tgatataaac ctcctcaaac attttactta gaggcaagga ttgtctaatt 1496 tcaattgtgc aagacatgtg ccttataatt atttttagct taaaattaaa cagattttgt 1556 aataatgtaa ctttgttaat aggtgcataa acactaatgc agtcaatttg aacaaaagaa 1616 gtgacataca caatataaat catatgtctt cacacgttgc ctatataatg agaageaget 1676 ctctgagggt tctgaaatca atgtggtccc tctcttgccc actaaacaaa gatggttgtt 1736 cggggtttgg gattgacact ggaggeagat agttgcaaag Ttgtetaag gttt ccctag 1796 ctgtatttag cctctgacta tattagtata caaagaggtc atgtggttga gaocaggtga 1856 atagtcacta tcagtgtgga gacaagcaca gcacacagac attttaggaa ggaaaggaac 1916 taegaaateg tgtgaaaatg ggttggaacc catcagtgat egeatattea ttgatgaggg 1976 tttgettgag atEigaaaatg gtggctcctt tctgtcttat ctcctagttt cttcaatgct 2036 taegeettgt tcttctcaag agaaagttgt aactctctgg tcttcatatg tccctgtgct 2096 ccttttaacc aaataaagag ttcttgtttc tgaagaa 2133 &lt; 210 &gt; 8 &lt; 211 &gt; 236 &lt;212&gt; PRT &lt;213> Homo sapiens &lt;400&gt; 8

Met Ala Tyr Phe His Glu Leu Ser Ser Arg Val Ser Phe Gin Glu Ala 15 10 15

Arg Leu Ala Cys Glu Ser Glu Gly Gly Val Leu Leu Ser Leu Glu Asn 20 25 30

Glu Ala Glu Gin Lys Leu lie Glu Ser Met Leu Gin Asn Leu Thr Lys 35 40 45

Pro Gly Thr Gly lie Ser Asp Gly Asp Phe Trp lie Gly Leu Trp Arg 50 55 60

Asn Gly Asp Gly Gin Thr Ser Gly Ala Cys Pro Asp Leu Tyr Gin Trp 65 70 75 80

Ser Asp Gly Ser Asn Ser Gin Tyr Arg Asn Trp Tyr Thr Asp Glu Pro 85 90 95

Ser Cys Gly Ser Glu Lys Cys Val Val Met Tyr His Gin Pro Thr Ala 100 105 110

Asn Pro Gly Leu Gly Gly Pro Tyr Leu Tyr Gin Trp Asn Asp Asp Arg 115 120 125

Cys Asn Met Lys His As Tyr lie Cys Lys Tyr Glu Pro Glu lie Asn 130 135 140

Pro Thr Ala Pro Val Glu Lys Pro Tyr Leu Thr Asn Gin Pro Gly Asp 145 150 155 160

Thr His Gin Asn Val Val Val Thr Glu Ala Val Lys Glu Glu Gin Lys 165 170 175

Leu Val Gin Thr Ser Leu His Cys Gly Phe Gin Arg Val Pro Glu Lys 180 185 190

Lys Val Ala Trp Lys Tyr Asn Asn Ser Leu Thr Trp Phe Gin Asn Phe 195 200 205

Val lie Leu Asp Leu Tyr Lys Glu Trp His Gin Asn Asn Ser Leu Glu 210 215 220

Trp Leu Glu lie Thr Lys Asp Leu Gin Asp Glu Leu 225 230 235 &lt;210&gt; 9 &lt;211&gt; 2033 &lt;212> DNA &lt;213> Homo sapiens &lt;220&gt;

&lt;221> CDS &lt;221>(106)(816) 142769-sequence table.doc 201021828 &lt;400&gt; 9 gcttgaagat ggcagatcgt gggacttctc agcctccata attgagtgag ccaattccct 60 ggccaaaagg tgtgttttgc tgacttcaag catccctgct acaaa atg gcc tac ttc 117

Met Ala Tyr Phe cat gaa ctg tcc age ega gtg age ttt cag gag gca ege ctg get tgt 165

His Glu Leu Ser Ser Arg Val Ser Phe Gin Glu Ala Arg Leu Ala Cys 5 10 15 20 gag agt gag gga gga gtc etc etc age ett gag aat gaa gca gaa cag 213

Glu Ser Glu Gly Gly Val Leu Leu Ser Leu Glu Asn Glu Ala Glu Gin 25 30 35 aag tta ata gag age atg ttg caa aac ctg aca aaa ccc ggg aca ggg 261

Lys Leu He Glu Ser Met Leu Gin Asn Leu Thr Lys Pro Gly Thr Gly 40 45 50 att tet gat ggt gat ttc tgg ata ggg ett tgg agg aat gga gat ggg 309 lie Ser Asp Gly Asp Phe Trp lie Gly Leu Trp Arg Asn Gly Asp Gly 55 60 65 caa aca tet ggt gcc tgc cca gat etc tac cag tgg tet gat gga age 357

Gin Thr Ser Gly Ala Cys Pro Asp Leu Tyr Gin Trp Ser Asp Gly Ser

70 75 80 aat tec cag tac ega aac tgg tac aca gat gaa cct tec tgc gga agt 405

Asn Ser Gin Tyr Arg Asn Trp Tyr Thr Asp Glu Pro Ser Cys Gly Ser 85 90 95 100 gaa aag tgt gtt gtg atg tat cac caa cca act gcc aat cct ggc ett 453

Glu Lys Cys Val Val Met Tyr His Gin Pro Thr Ala Asn Pro Gly Leu 105 110 115 ggg ggt ccc tac ett tac cag tgg aat gat gac agg tgt aac atg aag 501

Gly Gly Pro Tyr Leu Tyi- Gin Trp Asn Asp Asp Arg Cys Asn Met Lys 120 125 130 cac aat tat att tgc aag tat gaa cca gag att aat cca aca gcc cct 549

His Asn Tyr lie Cys Lys Tyr Glu Pro Glu lie Asn Pro Thr Ala Pro 135 140 145 gta gaa aag cct tat ett aca aat caa cca gga gac acc cat cag aat 597

Val Glu Lys Pro Tyr Leu Thr Asn Gin Pro Gly Asp Thr His Gin Asn 150 155 160 645 693 gtg gtt gtt act gaa gca gta aag gaa gaa caa aaa eta gtc caa acc

Val Val Val Thr Glu Ala Val Lys Glu Glu Gin Lys Leu Val Gin Thr 165 170 175 180 agt eta cac tgt gga ttt caa aga gta cca gaa aag aaa gtg gca tgg

Ser Leu His Cys Gly Phe Gin Arg Val Pro Glu Lys Lys Val Ala Trp 185 190 195 aag tat aat aac tea ttg act tgg ttc cag aat ttt gta att ctg gat 741

Lys Tyr Asn Asn Ser Leu Thr Trp Phe Gin Asn Phe Val lie Leu Asp 200 205 210 ctg tat aag gaa tgg cat cag aac aat age ttg gaa tgg ett gaa ate 789

Leu Tyr Lys Glu Trp His Gin Asn Asn Ser Leu Glu Trp Leu Glu lie 215 220 225 aca aag gat ctg caa gat gaa ctg taa gctccccctt gaggcaaata 836

Thr Lys Asp Leu Gin Asp Glu Leu * 230 235 ttaaagtaat ttttatatgt ctattatttc atttaaagaa taataatgga 896 gtgagacatg cttattttgc taaaggatgc acccaaactt caaacttcaa gcaaatgaaa 956 tggacaatgc agataaagtt gttatcaaca cgtcgggagt atgtgtgtta gaagcaattc 1016 cttttatttc tttcaccttt cataagttgt tatetagtea atgtaatgta tattgtattg 1076 aaatttacag tgtgcaaaag tattttacct ttgcataagt gtttgataaa aatgaactgt 1136 tetaatattt atttttatgg catctcattt ttcaatacat tatgctgtgc gctcttttga ttaaagaaac 1196 ttattactgt tgtcaactga attcacacac acacaaatat agtaccatag aaaaagtttg 1256 ttttctcgaa ataattcatc tttcagcttc tctgcttttg gtcaatgtct aggaaatctc 1316 142769 · sequence Listing .doc 201021828 ttcagaaata agaagctatt tcattaagtg tgatataaac ctcctcaaac attttactta 1376 gaggcaagga ttgtctaatt tcaattgtgc £ Lagacatgtg ccttataatt atttttagct 1436 taaaattaaa cagattttgt aataatgtaa ctttgttaat aggtgcataa acactaatgc 1496 agtcaatttg aacaaaagaa gtgacataca caatataaat Catatgtctt cacacgttgc 1556 ctatataatg agaagcagct ctctgagggt tctgaaatca atgtggtccc tctcttgccc 1616 act aaacaaa gatggttgtt cggggtttgg gattgacact ggaggcagat agttgcaaag 1676 ttagtctaag gtttc ^ ctag ctgtatttag cctctgacta tattagtata caaagaggtc 1736 atgtggttga gaccaggtga atagtcacta tcagtgtgga gacaagcaca gcacacagac 1796 attttaggaa ggaaaggaac tacgaaatcg tgtgaaaatg ggttggaacc catcagtgat 1856 cgcatattca ttgatgaggg tttgcttgag atagaaaatg gtggctcctt tctgtcttat 1916 ctcctagttt cttcaatgct tacgccttgt tcttctcaag agaaagttgt aactctctgg 1976 tcttcatatg tccctgtgct ccttttaacc aaataaagag ttcttgtttc tgaagaa 2033 &lt;210&gt; 10 &lt;211> 236 &lt;212&gt; PRT &lt;213> Homo sapiens &lt;400> 10

Met Ala Tyr Phe His Glu Leu Ser Ser Arg Val Ser Phe Gin Glu Ala 15 10 15

Arg Leu Ala Cys Glu Ser Glu Gly Gly Val Leu Leu Ser Leu Glu Asn 20 25 30

Glu Ala Glu Gin Lys Leu lie Glu Ser Met Leu Gin Asn Leu Thr Lys 35 40 45

Pro Gly Thr Gly lie Ser Asp Gly Asp Phe Trp lie Gly Leu Trp Arg 50 55 60

Asn Gly Asp Gly Gin Thr Ser Gly Ala Cys Pro Asp Leu Tyr Gin Trp 65 70 75 80

Ser Asp Gly Ser Asn Ser Gin Tyr Arg Asn Trp Tyr Thr Asp Glu Pro 85 90 95

Ser Cys Gly Ser Glu Lys Cys Val Val Met Tyr His Gin Pro Thr Ala 100 105 110

Asn Pro Gly Leu Gly Gly Pro Tyr Leu Tyr Gin Trp Asn Asp Asp Arg 115 120 125

Cys Asn Met Lys His As Tyr lie Cys Lys Tyr Glu Pro Glu lie Asn 130 135 140

Pro Thr Ala Pro Val Glu Lys Pro Tyr Leu Thr Asn Gin Pro Gly Asp 145 150 155 160

Thr His Gin Asn Val Val Val Thr Glu Ala Val Lys Glu Glu Gin Lys 165 170 175

Leu Val Gin Thr Ser Leu His Cys Gly Phe Gin Arg Val Pro Glu Lys 180 185 190

Lys Val Ala Trp Lys Tyr Asn Asn Ser Leu Thr Trp Phe Gin Asn Phe 195 200 205

Val lie Leu Asp Leu Tyr Lys Glu Trp His Gin Asn Asn Ser Leu Glu 210 215 220

Trp Leu Glu lie Thr Lys Asp Leu Gin Asp Glu Leu 225 230 235 &lt;210&gt; 11 &lt;211&gt; 649 &lt;212&gt; DNA &lt;213> Homo sapiens &lt;220&gt;221&gt; CDS &lt;222&gt; 39). . . (648) &lt;400&gt; 11 aatactttct gagagtcctg gacctcctgt gcaagaac atg aaa cac ctg tgg ttc 56

Met Lys His Leu Trp Phe 1 5 ttc etc etc ctg gtg gca get ccc aga tgg gtc ctg tcc cag gtg cag 104

Phe Leu Leu Leu Val Ala Ala Pro Arg Trp Val Leu Ser Gin Val Gin 10 15 20 ctg cag gag teg ggc cca gga ctg gtg aag cct teg gag acc ctg tcc 152

Leu Gin Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser 25 30 35 etc acc tgc act gtc tet ggt ggc tcc gtc age agt ggt ggt tac tac 200

Leu Thr Cys Thr Val Ser Gly Gly Ser Val Ser Ser Gly Gly Tyr Tyr -10· 142769 - Sequence Listing.doc 248 201021828 248 40 45 50

Tgg age tgg ate egg cag ccc cca ggg aag gga ctg gag tgg att ggg

Trp Ser Trp lie Arg Gin Pro Pro Gly Lys Gly Leu Glu Trp lie Gly 55 60 65 70 tat ate tat tac agt ggg ggc acc aac tac aac ccc tee etc aag agt

Tyr lie Tyr Tyr Ser Gly Gly Thr Asn Tyr Asn Pro Ser Leu Lys Ser 75 80 85 ega gtc acc ata tea gta gac aeg tee aag aac cag ttc tee ctg aag

Arg Val Thr lie Ser Val Asp Thr Ser Lys Asn Gin Phe Ser Leu Lys 90 95 100 ctg acc tet gtg acc get geg gac aeg gee gtg tat tac tgt geg aga

Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg 105 110 115 gag teg gga tat tgt act aat gtt gca tgc ttc cct gat get ttt gat

Glu Ser Gly Tyr Cys Thr Asn Val Ala Cys Phe Pro Asp Ala Phe Asp 120 125 130 ate tgg ggc caa ggg aca atg gtc acc gtg tet tea gee tee acc aag lie Trp Gly Gin Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys 135 140 145 150 ggc cca teg gtc ttc ccc ctg gca ccc tee tee aag age acc tet ggg

Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 155 160 165 ggc aca geg gee ctg ggc tgc ctg gtc aag gac tac ttc ccc gaa ccg Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro 170 175 180 gtg aeg gtg teg tgg aac tea ggc gee ctg acc age ggc gtg cac acc Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 185 190 195 ttc cca get gtc eta ca Phe Pro Ala Val Leu 200 &lt;210> 12 &lt;211&gt; 203 &lt;212> PRT &lt;213> Homo sapiens &lt;400> 12

Met Lys His Leu Τχ-p Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp 15 10 15

Val Leu Ser Gin Val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val Lys 20 25 30

Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Val 35 40 45

Ser Ser Gly Gly Tyr Tyr Trp Ser Trp lie Arg Gin Pro Pro Gly Lys 50 55 60

Gly Leu Glu Trp lie Gly Tyr lie Tyr Tyr Ser Gly Gly Thr Asn Tyr 65 70 75 80

Asn Pro Ser Leu Lys Ser Arg Val Thr lie Ser Val Asp Thr Ser Lys 85 90 95

Asn Gin Phe Ser Leu Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala 100 105 110

Val Tyi' Tyr Cys Ala Arg Glu Ser Gly Tyr Cys Thr Asn Val Ala Cys 115 120 125

Phe Pro Asp Ala Phe Asp lie Trp Gly Gin Gly Thr Met Val Thr Val 130 135 140

Ser Ser Ala Ser Thr Lys Gly Px*o Ser Val Phe Pro Leu Ala Pro Ser 145 150 155 160

Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys 165 170 175

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu 180 185 190

Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 195 200 142769 - Sequence Listing.doc -11- 296 344 392 440 488 536 584 632 649 201021828 &lt;210> 13 &lt;211> 884 &lt;212> DNA &lt;213 〉 Homo sapiens &lt;220&gt;

&lt;221> CDS &lt;222>(100). .. (81® &lt;400> 13 caccgcggtg gcggccgctc tagcccgact ggagcacgag gacactgaca tggactgatg gagtagaaag atcaggactc ctcagttcac cttctcaca atg agg etc cct get

Met Arg Leu Pro Ala cag etc ctg ggg ctg eta atg etc tgg gtc cca gga tee agt ggg gat

Gin Leu Leu Gly Leu Leu Met Leu Trp Val Pro Gly Ser Ser Gly Asp 10 15 20 gtt gtg atg act cag tet cca etc tee ctg ccc gtc acc ett gga cag

Val Val Met Thr Gin Ser Pro Leu Ser Leu Pro Val Thr Leu Gly Gin 25 30 35 ccg gee tee ate tee tgc agg tet agt caa age etc gta tac act gat

Pro Ala Ser lie Ser Cys Arg Ser Ser Gin Ser Leu Val Tyr Thr Asp 40 45 50 gga aac acc tee ttg aat tgg ttt cag cag agg cca ggc caa tet cca

GG ge ge eta att tat aag gtt tet aac tgg gac tet ggg gtc cca gac

Arg Arg Leu lie Tyr Lys Val Ser Asn Trp Asp Ser Gly Val Pro Asp 70 75 80 85 age ttc age ggc agt ggg tea ggc act gat ttc aca ctg aaa ate age

Ser Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie Ser 90 95 100 agg gtg gag get gaa aat gtt ggg gtt tat tac tgc atg caa ggt aca

Arg Val Glu Ala Glu Asn Val Gly Val Tyr Tyr Cys Met Gin Gly Thr 105 110 115 cac tgg cct ttc act ttc ggc gga ggg acc eiag gtg gag ate aaa ega

His Trp Pro Phe Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg 120 125 130 act gtg get gca cca tet gtc ttc ate ttc ccg cca tet gat gag cag

Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin 135 140 145 ttg aaa tet gga act gee tet gtt gtg tgc ctg ctg aat aac ttc tat

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 150 155 160 165 ccc aga gag gee aaa gta cag tgg aag gtg gat aac gee etc caa teg

Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser 170 175 180 ggt aac tee cag gag agt gtc aca gag cag gac age aag gac age acc

Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr 185 190 195 tac age etc age age acc ctg aeg ctg age aaa gca gac tac gag aaa

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 200 205 210 cac aaa gtc tac gee tgc gaa gtc acc cat cag ggc ctg age teg ccc

His Lys Val Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro 215 220 225 gtc aca aag age ttc aac agg gga gag tgt tagagggegg atcccccggg Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 12- 142769 - Sequence Listing.doc 201021828 230 235 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggg 884 &lt;210&gt; 14 &lt;211> 239 &lt;212> PRT &lt;213> Homo sapiens &lt;400> 14

Met Arg Leu Pro Ala Gin Leu Leu Gly Leu Leu Met Leu Trp Val Pro 15 10 15

Gly Ser Ser Gly Asp Val Val Met Thr Gin Ser Pro Leu Ser Leu Pro 20 25 30

Val Thr Leu Gly Gin Pro Ala Ser lie Ser Cys Arg Ser Ser Gin Ser 35 40 45

Leu Val Tyr Thr Asp Gly Asn Thr Ser Leu Asn Trp Phe Gin Gin Arg 50 55 60

Pro Gly Gin Ser Pro Arg Arg Leu lie Tyr Lys Val Ser Asn Trp Asp 65 70 75 80

Ser Gly Val Pro Asp Ser Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95

Thr Leu Lys lie Ser Arg Val Glu Ala Glu Asn Val Gly Val Tyr Tyr 100 105 110

Cys Met Gin Gly Thr His Trp Pro Phe Thr Phe Gly Gly Gly Thr Lys 115 120 125

Val Glu lie Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro 130 135 140

Pro Ser Asp Glu Gin Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp 165 170 175

Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp 180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gin 210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 &lt;210&gt; 15 &lt;211> 1016 &lt;212> DNA &lt;213> Homo sapiens &lt;220〉

&lt;221> CDS &lt;221> (178)...(899) &lt;400&gt; 15 cacacaggaa acagctatga ccatgattac gccaagcgcca caattaaccc tcactaaagg 60 gaacaaaagc tggagctcca ccgcggtggc ggccgctcta gcccggctgg agcacgagga 120 cactgacacg gactgaagga gtagaaagag ctacaacagg caggcagggg cagcaag atg 180

Met gtg ttg cag acc cag gtc ttc att tct ctg ttg etc tgg ate tet ggt 228

Val Leu Gin Thr Gin Val Phe lie Ser Leu Leu Leu Trp lie Ser Gly 5 10 15 gcc aac ggg gac ate gtg atg acc cag tct cca gac tcc ctg get gtg 276

Ala Asn Gly Asp lie Val Met Thr Gin Ser Pro Asp Ser Leu Ala Val 20 25 30 tct ctg ggc gag egg gcc acc ate aac tgc aag tcc age cag ggt gtt 324

Ser Leu Gly Glu Arg Ala Thr lie Asn Cys Lys Ser Ser Gin Gly Val 35 40 45 tta tac age tcc aac aat aag aac tac tta get tgg tac cag cag aaa 372

Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys 50 55 60 65 cca gga cag cca cct aag ctg etc att tac tgg gca tct acc egg gaa 420 -13· 142769-sequence table.doc 201021828

Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg Glu 70 75 80 tcc ggg gtc cct gac cga ttc agt ggc age ggg tet ggg aca gat ttc 468

Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95 act etc acc ate age age ctg cag get gaa gat gtg gca gtt tat ttc 516

Thr Leu Thr lie Ser Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Phe 100 105 110 tgt cag caa tat tat gtt agt ccg etc act ttc ggc gga ggg acc aag 564

Cys Gin Gin Tyr Tyr Val Ser Pro Leu Thr Phe Gly Gly Gly Thr Lys 115 120 125 gtg gag ate aaa cga act gtg get gca cca tet gtc ttc ate ttc ccg 612

Val Glu lie Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro 130 135 140 145 cca tet gat gag cag ttg aaa tet gga act gee tet gtt gtg tgc ctg 660

Pro Ser Asp Glu Gin Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 150 155 160 ctg aat aac ttc tat ccc aga gag gee aaa gta cag tgg aag gtg gat 708

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp 165 170 175 aac gee etc caa teg ggt aac tec cag gag agt gtc aca gag cag gac 756

Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp 180 185 190 age aag gac age acc tac age etc age age acc ctg aeg ctg age aaa 804

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 195 200 205 gca gac tac gag aaa cac aaa gtc tac gee tgc gaa gtc acc cat cag 852

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gin 210 215 220 225 ggc ctg age teg ccc gtc aca aag age ttc aac agg gga gag tgt ta 899

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 230 235 240 gagggeggat cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc 959 gagggggggc ccggtaccca attcgcccta tagtgagteg tattaegege gctcact 1016 &lt;210> 16 &lt;211&gt; 240 &lt;212> PRT &lt;213 〉智智&lt;400&gt; 16

Met Val Leu Gin Thr Gin Val Phe lie Ser Leu Leu Leu Trp lie Ser 15 10 15

Gly Ala Asn Gly Asp lie Val Met Thr Gin Ser Pro Asp Ser Leu Ala 20 25 30

Val Ser Leu Gly Glu Arg Ala Thr lie Asn Cys Lys Ser Ser Gin Gly 35 40 45

Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gin Gin 50 55 60

Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg 65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 85 90 95

Phe Thr Leu Thr lie Ser Ser Leu Gin Ala Glu Asp Val Ala Val Tyr 100 105 110

Phe Cys Gin Gin Tyr Tyr Val Ser Pro Leu Thr Phe Gly Gly Gly Thr 115 120 125

Lys Val Glu lie Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe 130 135 140

Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly Thr Ala Ser Val Val Cys 145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val 165 170 175

Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val Thr Glu Gin -14-

142769 · Sequence Listing.doc 201021828 180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser 195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His 210 215 220

Gin Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 240 &lt;210> 17 &lt;211&gt; 203 &lt;212&gt; PRT &lt;213> Homo sapiens &lt;400&gt;

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp 15 10 15

Val Leu Ser Gin Val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val Lys 20 25 30

Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Val 35 40 45

Ser Ser Gly Gly Tyr Tyr Trp Ser Trp lie Arg Gin Pro Pro Gly Lys 50 55 60

Gly Leu Glu Trp lie Gly Tyr He Tyr Tyr Ser Gly Gly Thr Asn Tyr 65 70 75 80

Asn Pro Ser Leu Lys Ser Arg Val Thr lie Ser Val Asp Thr Ser Lys 85 90 95

Asn Gin Phe Ser Leu Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala 100 105 110

Val Tyr Tyr Cys Ala Arg Glu Ser Gly Tyr Cys Thr Asn Val Ala Cys 115 120 125

Phe Pro Asp Ala Phe Asp lie Trp Gly Gin Gly Thr Met Val Thr Val 130 135 140

Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser 145 150 155 160

Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys 165 170 175

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu 180 185 190

Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 195 200 &lt;210&gt; 18 &lt;211> 239 &lt;212> PRT &lt;213> Homo sapiens &lt;400&gt; 18

Met Arg Leu Pro Ala Gin Leu Leu Gly Leu Leu Met Leu Trp Val Pro 15 10 15

Gly Ser Ser Gly Asp Val Val Met Thr Gin Ser Pro Leu Ser Leu Pro 20 25 30

Val Thr Leu Gly Gin Pro Ala Ser lie Ser Cys Arg Ser Ser Gin Ser 35 40 45

Leu Val Tyr Thr Asp Gly Asn Thr Ser Leu Asn Trp Phe Gin Gin Arg 50 55 60

Pro Gly Gin Ser Pro Arg Arg Leu lie Tyr Lys Val Ser Asn Trp Asp 65 70 75 80

Ser Gly Val Pro Asp Ser Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95

Thr Leu Lys lie Ser Arg Val Glu Ala Glu Asn Val Gly Val Tyr Tyr 100 105 110

Cys Met Gin Gly Thr His Trp Pro Phe Thr Phe Gly Gly Gly Thr Lys 115 120 125

Val Glu lie Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro 130 135 140

Pro Ser Asp Glu Gin Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp 165 170 175

Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp 180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 195 200 205 15- 142769 - Sequence Listing.doc 201021828

Ala Asp Tyr Glu Lys His Lys Val T-yr Ala Cys Glu Val Thr His Gin 210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 &lt;210> 19 &lt;211&gt; 240 &lt;212> PRT &lt;213> Homo sapiens &lt;400&gt;

Met Val Leu Gin Thr Gin Val Phe He Ser Leu Leu Leu Trp lie Ser 15 10 15

Gly Ala Asn Gly Asp lie Val Met Thr Gin Ser Pro Asp Ser Leu Ala 20 25 30

Val Ser Leu Gly Glu Arg Ala Thr I He Asn Cys Lys Ser Ser Gin Gly 35 40 45

Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gin Gin 50 55 60

Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg 65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 85 90 95

Phe Thr Leu Thr lie Ser Ser Leu GUn Ala Glu Asp Val Ala Val Tyr 100 105 110

Phe Cys Gin Gin Tyr Tyr Val Ser Paro Leu Thr Phe Gly Gly Gly Thr 115 120 125

Lys Val Glu lie Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe 130 135 140

Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly Thr Ala Ser Val Val Cys 145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg GIlu Ala Lys Val Gin Trp Lys Val 165 170 175

Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val Thr Glu Gin 180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser L^u Ser Ser Thr Leu Thr Leu Ser 195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His 210 215 220

Gin Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 240 16· 142769 - Sequence Listing 』0 (:

Claims (1)

201021828 VII. Patent Application Range: 1. An isolated monoclonal antibody or fragment thereof comprising an antigen binding site that specifically binds to a 58P1D12 protein comprising the amino acid sequence of SEQ ID NO: 2 and wherein the single The antibody of the strain comprises SEq ID NO: residues V to 20 to 146 of Vh region and residues 21 to 134 of VL of SEQ ID NO: 19. 2. The antibody or fragment of claim 1, wherein the antibody comprises SEQ. ID NO: The light chain sequence shown in 21 to 240 of 19 and the heavy chain sequence comprising the sequence shown in 20 to 203 of SEQ.® ID NO: 17. 3. The antibody or fragment of claim 1, wherein the amino acid sequence of the VH and VL regions of the antibody is the same as the antibody produced by the fusion of the designated a.t.c.C. accession number 9404. 4. The antibody or fragment of claim 1, wherein the fragment is a Fab, F(ab')2, Fv or Sfv fragment. 5. The antibody or fragment of claim 1, which is prepared recombinantly. 6. The antibody or fragment of claim 5, wherein the recombinant protein comprises the anti-scorpion binding region. 7. The antibody or fragment of claim 1 wherein the anti-system is coupled to a detectable target, toxin, therapeutic agent or chemotherapeutic agent. 8. The antibody or fragment of claim 7, wherein the detectable marker is a radioisotope, a metal chelator, an enzyme, a fluorescent compound, a bioluminescent compound, or a chemiluminescent compound. 9. The antibody or fragment of claim 8, wherein the radioisotope comprises 212Bi &gt;, 3, I '131In ' 9〇Y ' , 86Re &gt; 21, At ' 125I ^ 188Re &gt; 142769.doc 201021828 153Sm, 213Bi , 32P or Lu. 10. The antibody or fragment of claim 7, wherein the toxin comprises ricin, anthraquinone A chain, doxorubicin, dauno rubicin, maytansinoid ), taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincri stine, Vinblastine, colchicine 'dihydroxy anthracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin ; PE) A, PE40, abrin toxin (abrin), abrin toxin A chain, modeccin A chain, alpha sarcin, gelonin, mitigellin ), retstrlctocin, phenomycin, enomycin, curicin, crotin, calicheamicin, saponaria (sapaonaria officinalis) inhibitor, Corticosteroids, Orlista; auristatin, auromycin, ge, silk, combestatin, duocarmycin, dolostatin , ccl065 or cisplatin. 11. The antibody or fragment of claim 1, wherein the antigen binding site specifically binds to an epitope within the amino acid sequence of SEQ ID NO: 2. 12. A fertile genetic animal which produces a monoclonal antibody as claimed in claim 1. 13. A fusion tumor which produces a monoclonal antibody as claimed in claim 1. 142769.doc 201021828 14. A polynucleotide encoding a light or heavy chain of an antibody as claimed. 15. A vector comprising the polynucleotide of claim 14. 16. The vector of claim 15 which is a single strand comprising a variable domain of a heavy chain and a light chain. 17. A cell transfected with a vector of claim 15. 18. The cell of claim 17, wherein the cell line is transfected with a vector comprising a polynucleotide encoding the light chain of the antibody of claim 1 and a polynucleotide encoding the heavy chain of the antibody of claim 1. Or a vector comprising a polynucleotide comprising a light chain of an antibody of claim 1 and a vector comprising a polynucleotide encoding a heavy chain of an antibody of the claim 转. 19. A method for producing a light chain variable region comprising: 21 to 134 of *SEQ ID NO: 19 and a heavy chain variable region as set forth in 19 to 146 of SEQ. ID NO: 17. A method of antibody or fragment of a sequence, the method comprising: i) cultivating the cell of claim 17 under conditions which promote expression of the antibody or fragment, and π) isolating the antibody or fragment from the cells, thereby preparing the antibody Or a fragment. 20. The method of claim 19, wherein the antibody comprises a light chain sequence as shown in 21 to 240 of SEq m N〇:i9* and comprises 19th to 203th as in SEq. iD n〇17 The heavy chain sequence of the sequence shown. 21. A pharmaceutical composition comprising an antibody or fragment of claim 1 in a human unit dosage form. 22. A method for detecting the presence of a 58P1D12 protein in a biological sample, the assay comprising contacting the sample with an antibody of claim 1 and detecting the amino acid sequence comprising SEQ ID NO: 2 in the sample The knot of protein 142769.doc 201021828 Synergy. 23. A method of inhibiting cell growth in a subject 581 &gt; 1 〇 12, comprising: administering to the individual a vector encoding a single-chain monoclonal antibody comprising a specific binding to the inclusion The heavy chain and light chain variable domain of the monoclonal antibody of the 58P1D12 protein of the amino acid sequence of ID NO: 2 such that the vector can transmit the single-chain monoclonal antibody coding sequence to the cancer cell, and The encoded single chain antibody is expressed in the cell. A method of delivering a cytotoxic or diagnostic agent to a cell expressing a 58p丨〇丨2 protein, the method comprising: providing a cytotoxic or diagnostic agent that binds to an antibody or fragment of claim 1 to form An antibody agent or fragment agent conjugate; and exposing the cell to the antibody agent or fragment agent conjugate, such that the cytotoxic or diagnostic agent is delivered to the cell by the antibody or fragment thereof comprising SEQ ID NO: Amino acid sequence of this protein. The method of claim 24, wherein the cytotoxic agent or the diagnostic agent is selected to be a group of the target, the toxin, and the therapeutic agent. 26. The method of claim 25, wherein the detectable marker is a radioisotope, a metal chelator, an enzyme, a fluorescent compound, a bioluminescent compound, or a chemiluminescent compound. 27. The method of claim 26, wherein the radioisotope comprises 2!2Bi, 31I, 131In '90Y, 186Re, 2&quot; At, %, 18&amp;, |53sm, 213Bi, 32P or Lu. The method of claim 25, wherein the toxin comprises ricin, ricin 142769.doc 201021828 A chain, doxorubicin, daunorubicin, maytansinol, paclitaxel, ethidium bromide, Mitomycin, Etoposide, Tenopore, Vincristine, Vinblastine, Colchicine, Dihydroxy Anthrax Dione, Actinomycin, Diphtheria, Pseudomonas aeruginosa Exotoxin (PE) A , PE40, scorpion toxin, Acacia toxic toxin A chain, Modicin A chain, alpha sputum, leucovorin, mitogen, quercetin, phenolic acid, indomycin, jatropha, Crotonin, calicheamicin, saponin inhibitor, glucocorticoid, orlistatin chlortetracycline, guanidine, guanidine, compostatin, dokanicin, dolostatin, ccl〇 65 or cisplatin. 29. A method for detecting a 58P1D12 protein in a biological sample, the method comprising the steps of: providing the biological sample and a control sample; and causing the biological sample and the control sample to specifically bind to the 58P1D12 protein as claimed in claim i The antibody is contacted, wherein the protein comprises the amino acid sequence of SEQ ID NO: 2; and the amount of the substance and the complex of the 58p丨D丄2 protein and the antibody in the control sample and the control sample. 30. The method of claim 29, further comprising: collecting the biological sample and the control sample from a patient having or suspected of having the cancer listed in Table I. 31. A composition comprising: 58P1D12 siRNA (double stranded RNA) corresponding to a nucleic acid encoding a protein comprising an amino acid sequence of SEq id NO: 2, wherein the subsequence has 19, 2, 21, 22, 23, 24 or 25 sequences of RNA nucleotide lengths that are complementary and non-complementary to at least one of the I42769.doc 201021828 portions of the mRNA coding sequence. 3 2. A method of identifying a molecule that modulates cell proliferation, the method comprising: (a) introducing a molecule into a system comprising a nucleic acid, the nucleic acid comprising: a nucleotide sequence selected from the group consisting of: (i) a nucleotide sequence of SEQ ID NO: 1; (Π) a nucleotide sequence encoding a polypeptide comprising residues 19 to 134 of the amino acid sequence of SEQ ID NO: I9; (in) coding with SEQ ID NO: 19 amino acid sequence 栌 residues 19 to 134 have 90 ° /. Or a nucleotide sequence of more than 90% of the consensus polypeptide; and (iv) a fragment of the nucleotide sequence of (i), (ii) or (iii); or introducing a test molecule to comprise (i), Or a system of the protein encoded by the acid sequence of ii), (iii) or (iv); and (b) determining the presence or absence of an interaction between the molecule and the nucleotide sequence or protein genus, The molecule interacts with the nucleotide sequence or protein to identify the molecule as a molecule that regulates cell proliferation. 33. The method of claim 32, wherein the system is an in vivo system. 34. The method of claim 32, wherein the system is an in vitro system. 35. The method of claim 32, wherein the molecule comprises an antibody or antibody fragment that specifically binds to the white matter encoded by the nucleotide sequence of (1), GO, (iii) or (iv). In the method of claim 32, wherein the molecule is a composition, the aged composition comprises 58P1D12 siRNA (double stranded RNA) corresponding to a nucleic acid encoding a protein comprising the amino acid sequence of SEQ ID NO: 2 or a malaria sequence. In 142769, doc 201021828, the subsequence has 19, 20, 21, 22, 23, 24 or 25 contiguous RNA nucleotide lengths and contains sequences that are complementary and non-complementary to at least a portion of the mRNA coding sequence. 37. A method of treating cancer in a subject' which comprises administering to a subject diagnosed with cancer a molecule identified by the method of claim 32, whereby the molecule inhibits or delays cancer in the individual. 3. The method of claim 37, wherein the cancer is a cancer listed in Table j. 39. A method of identifying a therapeutic agent for treating cancer as listed in Table I, the method (a) introducing a molecule into a system comprising a nucleic acid comprising a nucleotide sequence selected from the group consisting of: (i a nucleotide sequence of SEQ ID NO: 1; (11) a nucleotide sequence encoding a polypeptide comprising residues 19 to 134 of the amino acid sequence of SEQ ID NO: 19; (iii) encoding with SEQ ID NO: a nucleotide sequence of a polypeptide having a 90% or greater homology of residues 19 to 134 in the amino acid sequence of 19; and (iv) a nucleotide sequence of (i), (ii) or (in) a fragment; or a system for introducing a test molecule to a protein comprising a nucleotide sequence encoded by (i), (ii), (iii) or (iv); and (b) determining between the molecule and the nucleoside The presence or absence of an acid sequence or interaction between proteins, whereby an interaction between the molecule and the nucleotide sequence or protein identifies the molecule as a therapeutic agent for the treatment of tissue cancer as listed in Table I. 142769.doc 201021828 40. 41. 42. 43. 44. 45. 46. 47. 48. The method of claim 39, wherein the system is alive Outside the system. The method of claim 39, wherein the system is an in vivo system. The method of claim 39, wherein the molecule comprises an antibody or antibody fragment that specifically binds to the protein encoded by the nucleotide sequence of (1), (ii), (iii) or (iv). The method of claim 39, wherein the molecule is a composition comprising a 58P1D12 siRNA (double stranded RNA) corresponding to a nucleic acid encoding a protein comprising the amino acid sequence of SEQ ID NO: 2 or a subsequence thereof, Wherein the subsequence has 19, 2, 21, 22, 23, 24 or 25 contiguous RNA nucleotide lengths and comprises a sequence that is complementary and non-complementary to at least a portion of the mRNA coding sequence. A method of treating cancer in a subject comprising administering to a subject diagnosed with cancer a molecule identified by the method of claim 39, whereby the molecule inhibits or delays cancer in the individual. The method of claim 44, wherein the cancer is a cancer listed in Table I. A method for reducing tumor growth in a mammal comprising the use of an effective amount of a monoclonal antibody, such as a monoclonal antibody, which is specific for binding to a protein comprising an amino acid sequence of SEq ID N〇: 2, and a radiotherapy method The mammal. A method of reducing tumor growth in a mammal comprising treating with a combination of a monoclonal antibody and a chemotherapeutic agent, as claimed, in an amount effective to bind to a protein comprising the amino acid sequence of SEQ ID NO: 2. Hermitian. - A method of reducing swelling and growth of a mammal comprising the use of an effective amount of 142769.doc 201021828 to specifically bind to a protein comprising the amino acid sequence of SEQ ID NO: 2, such as the request term α I The mammalian antibody is treated with a combination of an early strain antibody and a pharmaceutical or biologically active therapy. 49. A method for identifying 58P1D12 peptone, 丨8-7, a sputum small molecule conjugate, the method comprising: (1) providing an array of a plurality of small molecule compounds, wherein the small molecule array is capable of binding to SEQ ID The 58P1D12 protein of the amino acid sequence of Ν〇·2; (2) contacting the array with the protein of the · · and (3) identifying the small molecule collocation by (reciprocal) Things. 142769.doc