CN109678778A - A kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent - Google Patents
A kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent Download PDFInfo
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- CN109678778A CN109678778A CN201811618531.1A CN201811618531A CN109678778A CN 109678778 A CN109678778 A CN 109678778A CN 201811618531 A CN201811618531 A CN 201811618531A CN 109678778 A CN109678778 A CN 109678778A
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- Prior art keywords
- crosslinking agent
- functional protein
- protein crosslinking
- purification
- isodigeranyl functional
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/44—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members
- C07D207/444—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5
- C07D207/448—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. maleimide
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to purification & isolation technical field, especially a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent includes the following steps；Step 1: ingredient: including major ingredient according to weight percent: dimaleoyl imino alkyl acid 40-50%, methylene chloride 27-43%, thionyl chloride 5-10%, p-nitrophenol 5-10%, sodium carbonate 4-12%, Step 2: preparation: dimaleoyl imino alkyl acid and methylene chloride and thionyl chloride being added in reaction kettle according to aforementioned proportion and are stirred 2-3h, Step 3: synthetic reaction；Step 4: purification, isodigeranyl functional protein crosslinking agent synthetic route yield prepared by the present invention is higher, it is not related to the use of expensive reagent, it is at low cost, it is easy to industrialized production, nanometer membrane separation technique is used, the impurity such as small molecule and inorganic salts have been filtered out, the quality of isodigeranyl functional protein crosslinking agent is improved, product purity is increased to 99% or more from 95%.
Description
Technical field
The present invention relates to purification & isolation technical field more particularly to a kind of purification films of isodigeranyl functional protein crosslinking agent point
Separating process.
Background technique
Protein cross agent is small molecule compound, have 2 or more for specific groups (- NH2 ,-
COOH ,-HS etc.) reactive terminal, can be coupled respectively with 2 or more molecule, so that these molecules be made to be incorporated in
Together, people generally use glutaraldehyde to connect antibody and indicator (such as enzyme), but its as protein cross agent when the seventies
The disadvantage is that more selecting the crosslinking of specificity the eighties since crosslinked group is random, mixed and disorderly polymer easy to form
Agent, such as NHS(have obtained more widely answering in life science for-COOH) and maleimide (being directed to-HS)
With, it dexterously can be in protein interaction research with crosslinking agent, immunology, the fields such as treatment of cancer obtain unexpected
Harvest be especially widely used in the synthesis of antibody coupling drug at present in life science；
Traditional isodigeranyl functional protein crosslinking agent organic reaction post-processing relies primarily on the methods of distillation, recrystallization, few
Effect is preferable when quantitative response, but there is the disadvantages of complicated for operation, purification degree is not high, loss is larger when industrialization, so being used for
Drug effect after synthetic antibody coupling drug is had a greatly reduced quality.
Summary of the invention
The purpose of the present invention is to provide a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent, on solving
The problem of proposing in background technique is stated, present invention employs nanometer membrane separation techniques, the impurity such as small molecule and inorganic salts have been filtered out,
Improve the quality of isodigeranyl functional protein crosslinking agent.
To achieve the goals above, present invention employs following technical solutions:
The purification membrane separation process for designing a kind of isodigeranyl functional protein crosslinking agent, includes the following steps；
Step 1: ingredient: including major ingredient according to weight percent: dimaleoyl imino alkyl acid 40-50%, methylene chloride 27-
43%, thionyl chloride 5-10%, p-nitrophenol 5-10%, sodium carbonate 4-12%.
Step 2: preparation: dimaleoyl imino alkyl acid and methylene chloride and thionyl chloride being added according to aforementioned proportion
It is stirred 2-3h in reaction kettle, reaction kettle is then warming up to 25 DEG C -30 DEG C and carries out reaction 3-8h；
Step 3: synthetic reaction: it is equal that p-nitrophenol being added to the solvent obtained in reaction kettle with step 1 according to aforementioned proportion
Even mixing, backward reaction kettle in be added sodium carbonate carry out condensation reaction 3-8h obtain isodigeranyl functional protein crosslinking agent；
Step 4: purification:
S1, it solvent that step 3 obtains is added in compensating cylinder stores；
S2, it the solvent in compensating cylinder is pipelined in membrane module by the pressurization of charging pump with certain flow velocity carries out
UF membrane；
S3, less than retaining molecular weight material permeance membrane module formed penetrating fluid be discharged, greater than the object of retaining molecular weight
Concentrate after matter is then retained by membrane module flows back to compensating cylinder through pressure gauge and regulating valve.
Preferably, in step 3, after the completion of the synthetic reaction further include: concentration removes excessive thionyl chloride.
Preferably, in step 4, the pipeline between the charging pump and membrane module is spirally connected flowmeter.
Preferably, the left side top of the compensating cylinder is welded with spiral transferring hopper.
A kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent proposed by the present invention, beneficial effect are:
1, isodigeranyl functional protein crosslinking agent synthetic route yield prepared by the present invention is higher, is not related to the use of expensive reagent,
It is at low cost, it is easy to industrialized production；
2, present invention employs nanometer membrane separation techniques, have filtered out the impurity such as small molecule and inorganic salts, improve isodigeranyl function egg
The quality of white matter crosslinking agent, product purity are increased to 99% or more from 95%；
3, the application that the promotion for the product quality that the present invention purifies has pushed it in terms of biological medicine, can be improved applicable model
It encloses, meets the demand in terms of biological medicine.
Detailed description of the invention
Fig. 1 is the flowage structure figure that the present invention purifies.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1, referring to Fig.1, a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent, including following step
Suddenly；
Step 1: ingredient: including major ingredient according to weight percent: dimaleoyl imino alkyl acid 45%, methylene chloride 35%, chlorination
Sulfoxide 8%, p-nitrophenol 5%, sodium carbonate 7%.
Step 2: preparation: dimaleoyl imino alkyl acid and methylene chloride and thionyl chloride being added according to aforementioned proportion
It is stirred 2h in reaction kettle, reaction kettle is then warming up to 25 DEG C -30 DEG C and carries out reaction 5h；
Step 3: synthetic reaction: it is equal that p-nitrophenol being added to the solvent obtained in reaction kettle with step 1 according to aforementioned proportion
Even mixing, backward reaction kettle in be added sodium carbonate carry out condensation reaction 6h obtain isodigeranyl functional protein crosslinking agent, synthesize
After the reaction was completed further include: concentration removes excessive thionyl chloride；
Step 4: purification:
S1, it solvent that step 3 obtains is added in compensating cylinder 1 stores, the left side top of compensating cylinder 1 is welded with spiral transferring hopper
7；
S2, the solvent in compensating cylinder 1 is pipelined in membrane module 3 by the pressurization of charging pump 2 with certain flow velocity
UF membrane is carried out, the pipeline between charging pump 2 and membrane module 3 is spirally connected flowmeter 6；
S3, less than retaining molecular weight material permeance membrane module 3 formed penetrating fluid be discharged, greater than the object of retaining molecular weight
Concentrate after matter is then retained by membrane module 3 flows back to compensating cylinder 1 through pressure gauge 4 and regulating valve 5.
Embodiment 2, referring to Fig.1, a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent, including following step
Suddenly；
Step 1: ingredient: including major ingredient according to weight percent: dimaleoyl imino alkyl acid 50%, methylene chloride 35%, chlorination
Sulfoxide 5%, p-nitrophenol 5%, sodium carbonate 5%.
Step 2: preparation: dimaleoyl imino alkyl acid and methylene chloride and thionyl chloride being added according to aforementioned proportion
It is stirred 3h in reaction kettle, reaction kettle is then warming up to 25 DEG C -30 DEG C and carries out reaction 7h；
Step 3: synthetic reaction: it is equal that p-nitrophenol being added to the solvent obtained in reaction kettle with step 1 according to aforementioned proportion
Even mixing, backward reaction kettle in be added sodium carbonate carry out condensation reaction 5h obtain isodigeranyl functional protein crosslinking agent, synthesize
After the reaction was completed further include: concentration removes excessive thionyl chloride；
Step 4: purification:
S1, it solvent that step 3 obtains is added in compensating cylinder 1 stores, the left side top of compensating cylinder 1 is welded with spiral transferring hopper
7；
S2, the solvent in compensating cylinder 1 is pipelined in membrane module 3 by the pressurization of charging pump 2 with certain flow velocity
UF membrane is carried out, the pipeline between charging pump 2 and membrane module 3 is spirally connected flowmeter 6；
S3, less than retaining molecular weight material permeance membrane module 3 formed penetrating fluid be discharged, greater than the object of retaining molecular weight
Concentrate after matter is then retained by membrane module 3 flows back to compensating cylinder 1 through pressure gauge 4 and regulating valve 5.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (4)
1. a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent, which is characterized in that include the following steps；
Step 1: ingredient: including major ingredient according to weight percent: dimaleoyl imino alkyl acid 40-50%, methylene chloride 27-
43%, thionyl chloride 5-10%, p-nitrophenol 5-10%, sodium carbonate 4-12%；
Step 2: preparation: reacting dimaleoyl imino alkyl acid with methylene chloride and thionyl chloride addition according to aforementioned proportion
It is stirred 2-3h in kettle, reaction kettle is then warming up to 25 DEG C -30 DEG C and carries out reaction 3-8h；
Step 3: synthetic reaction: it is equal that p-nitrophenol being added to the solvent obtained in reaction kettle with step 1 according to aforementioned proportion
Even mixing, backward reaction kettle in be added sodium carbonate carry out condensation reaction 3-8h obtain isodigeranyl functional protein crosslinking agent；
Step 4: purification:
Storage in compensating cylinder (1) is added in S1, the solvent for obtaining step 3；
S2, the solvent in compensating cylinder (1) is pipelined to membrane module by the pressurization of charging pump (2) with certain flow velocity
(3) UF membrane is carried out in；
S3, less than retaining molecular weight material permeance membrane module (3) formed penetrating fluid be discharged, greater than retaining molecular weight
Substance then flows back to compensating cylinder (1) through pressure gauge (4) and regulating valve (5) by the concentrate after membrane module (3) retention.
2. a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent according to claim 1, it is characterised in that:
In step 3, after the completion of the synthetic reaction further include: concentration removes excessive thionyl chloride.
3. a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent according to claim 1, it is characterised in that:
In step 4, the pipeline between the charging pump (2) and membrane module (3) is spirally connected flowmeter (6).
4. a kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent according to claim 1, it is characterised in that:
The left side top of the compensating cylinder (1) is welded with spiral transferring hopper (7).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811618531.1A CN109678778A (en) | 2018-12-28 | 2018-12-28 | A kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent |
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| CN201811618531.1A CN109678778A (en) | 2018-12-28 | 2018-12-28 | A kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent |
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| CN201811618531.1A Pending CN109678778A (en) | 2018-12-28 | 2018-12-28 | A kind of purification membrane separation process of isodigeranyl functional protein crosslinking agent |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4980482A (en) * | 1988-01-19 | 1990-12-25 | The Dow Chemical Company | Process for the preparation of N-maleoyl activated esters of amino acids |
| US20030060441A1 (en) * | 2001-07-23 | 2003-03-27 | John-Stephen Taylor | Nucleic acid triggered catalytic drug and probe release |
| CN105037237A (en) * | 2015-06-12 | 2015-11-11 | 苏州昊帆生物科技有限公司 | Method for synthesizing N-maleimidoalkyl acid and succinimido ester thereof |
-
2018
- 2018-12-28 CN CN201811618531.1A patent/CN109678778A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4980482A (en) * | 1988-01-19 | 1990-12-25 | The Dow Chemical Company | Process for the preparation of N-maleoyl activated esters of amino acids |
| US20030060441A1 (en) * | 2001-07-23 | 2003-03-27 | John-Stephen Taylor | Nucleic acid triggered catalytic drug and probe release |
| CN105037237A (en) * | 2015-06-12 | 2015-11-11 | 苏州昊帆生物科技有限公司 | Method for synthesizing N-maleimidoalkyl acid and succinimido ester thereof |
Non-Patent Citations (1)
| Title |
|---|
| 少学时等: "《化工原理》", 31 August 2008 * |
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Application publication date: 20190426 |