JPH01233229A - Antibody and tooth decay preventive containing the same - Google Patents

Abstract

PURPOSE:To obtain an antibody which can be used as an active ingredient for preventing tooth decay, because it has satisfactory action to inhibit S. mutans from adhering to tooth surfaces, by preparing from eggs produced by hens immunized with a water-soluble glucan synthetase produced by S. mutans. CONSTITUTION:Streptococcus mutans of c, e or f serum type is cultured, as a pathogenic bacterium, in a medium containing at least glucose to obtain a water-soluble glucan synthetase (GFT-S) from the cell bodies. Hens are immunized using the enzyme as an antigen, eggs are collected from hens 1 or more months after immunization to prepare antiGFT-S antibody. High antibody values are kept for about 3 months. Since only IgG is transferred into the yolk, only IgG fraction can be specifically isolated through simple operation, namely yolk separation.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to the use of Stretococcus mutans, a pathogenic bacterium that induces portal erosion.
The present invention relates to an antibody that has an inhibitory activity against water-soluble glucan synthase (glucosyltransferase, abbreviated as GTF-3) produced by U.sub.tans), and a dental caries prevention agent containing the antibody as an active ingredient.

[Conventional technology]

It is already known that the process of attachment of S3,5utans to the tooth surface plays an important role in the development of portal caries, and it is known that the process of attachment of S0mutans to the tooth surface is controlled. Various attempts have been made to prevent portal erosion.

For example, a method for preventing portal erosion using an antibody having immunoactivity against S. mutans is known, and British Patent No. 1505513 describes
Breast milk obtained from cows immunized with S. muta
The method used to control the intraoral fixation of NS was also disclosed in Japanese Patent Application Laid-open No.
Publication No. 38327 discloses that antiserum and/or milk obtained by immunizing a mammal with cell walls obtained from culture of S1 trans, and a glucosyltransferase inhibitor, protease, and dextranase. A preventive method for preventing diarrhea using a combination of one or more synergists selected from the group has been described.

[Problem to be solved by the invention]

However, the preventive effect of conventional antibodies having immunoactivity against S. mutans is not sufficient, and conventional antibodies are prepared from the milk of immunized mammals or their antisera. , they lack mass productivity, have high production costs, and are often impractical.

The present inventors conducted various studies in order to solve the problems in the conventional technology, and as a result, the present inventors developed a method for preparing antibodies using chickens, which has the physicochemical properties described below.
A novel finding is that by using S. tautans water-soluble glucan synthase in combination as an antigen, it is possible to prepare an antibody that has a sufficient inhibitory effect on the adhesion of S. tautans to the tooth surface and does not have the above-mentioned drawbacks. Having obtained this knowledge, we have completed the present invention.

In particular, when preparing antibodies using chickens, it is not always possible to obtain a sufficient amount of antibodies that specifically react with the immunized antigen. For example, when immunizing with a virus antigen, a sufficient antibody titer may not be obtained. Ta.

Therefore, when immunizing chickens, it is necessary to carefully examine the antigen and make sure to obtain one with a high antibody titer.

Therefore, this is the first water-soluble glucan produced by S. mutans.

The object of the present invention is to exhibit immunoactivity against S. mutans, have a sufficient inhibitory effect on the adhesion of S. The object of the present invention is to provide an antibody that is useful as an active ingredient in a dental caries preventive agent, and a dental caries preventive agent containing the antibody.

[Means to solve the problem]

The antibody of the present invention having inhibitory activity against water-soluble glucan synthase (hereinafter abbreviated as GTF-3) produced by S, +mutans is an antibody of the present invention that has inhibitory activity against water-soluble glucan synthase (hereinafter abbreviated as GTF-3) produced by S, +mutans whose serotype is c, e, or f and has the following physicochemical characteristics. It can be obtained as immunoglobulin prepared from eggs produced by chickens immunized with GTF-5 produced by GTF-5.

0 action and substrate specificity Acts on sucrose to synthesize water-soluble glucan.

(2) Molecular weight 5DS - The molecular weight measured by polyacrylamide gel electrophoresis is 150 to 165 Daltons.

(2) Immunogenicity Acts as an immunogen in animals and can generate specific antibodies against the enzyme.

In addition, the above 5DS-polyacrylamide gel electrophoresis (
SO5-PAGE) was performed according to the following procedure.

5DS-PAGE operating conditions; 5DS-PAGE is performed by the method of Laea++li et al.

That is, each of the crude or purified samples described below was added to 2% S.
O3, 62.5 mM Tris-HCl buffer (pH 6) containing 5% 2-mercaptoethanol and 20% glycerol.
, 8) for 3 minutes at 100°C.

Electrophoresis was performed in a 7.5% separation gel containing 0.1% SDS and a 4% concentration gel at room temperature for 2 hours at 10 mA. Ferritin was used as a molecular weight marker. , 220 k Daltons), phosphorylase 1) (phosphorylaseb.

94 Dalton), bovine serum albumin (bovines)
erum albt++wins 67 k daltons), catalase (catalase, 60 k daltons), ovalbumin (ovalbumin s 43
k Dalton) and lactate dehydrogenase (Iactate
dehydrogenase, 36 k Daltons)
(both manufactured by Pharmacia), and the band can be detected by staining with Coomassie brilliant blue (CBB).

As GTF-3 used to obtain the antibody of the present invention (hereinafter referred to as anti-GTF-3), any enzyme having the above characteristics can be used. For example, S, +eutans can be prepared as follows. can be obtained by purification from the culture supernatant of

That is, S. mutans having serotypes c, e, or f are cultured in an appropriate medium and purified from the culture supernatant.

The type C seedlings used here include S, mutansI
Known and easily accessible bacteria such as ngbritt strain, M78148 strain, and 10449 strain can be used.
For example, S, mutan SMT8148 strain + In
gbritt strain was obtained from Osaka University School of Dentistry, and strain 10449 was obtained from the National Collection of Typical Cultures.
pe Cultures (NCTC)). Also, for e-type closure and f-type closure, known strains may be similarly obtained and used.

Further, as a medium, a medium containing at least glucose can be used, for example, TTY medium (Trypticase
, TryptoseSYeast extract
complex medium), BHI (Brain 1lea
rt Infusion medium, FMC medium, etc. can be used.

In addition, the culture temperature may be within a range suitable for bacterial cell growth and GTF-3 production, but in terms of good bacterial cell growth and GTF-3 production, it is usually around 37°C. It is good to say.

Further, the culture time varies depending on culture conditions such as culture temperature and type of medium, but may be determined by selecting the time when the optimum yield of GTF-3 is reached, and may be about 18 to 24 hours.

Further, other culture conditions may be appropriately selected from the above viewpoint.

Next, GTF-3 is extracted from the bacterial cells.

GTF-S of S, l1lutans is 5ato,
S, et al.'s method (Sato, S, + et al.
Carbohydr, Re5-+ 134+ 293
-304 (1984)), inoculate BHI dialysis medium with M. mutans, remove the bacterial cells by centrifugation after growth, add ammonium sulfate to the supernatant, and precipitate the 50% ammonium sulfate fraction with Histidine-HCI.
After dialysis and chromatofocusing column chromatography, a purified sample can be obtained by performing hydroxyapatite column chromatography.

As an antigen for obtaining antibodies, the bacterial cell culture supernatant or ammonium sulfate concentrated sample obtained by the above procedure, a crudely purified sample after chromatofocusing, or GT as a purified sample can be used.
F-3 etc. can be used.

Next, the method for preparing the anti-GTF-3 antibody of the present invention will be described in detail.

The anti-GTF-3 antibody of the present invention can be prepared from eggs of chickens immunized with the above-mentioned GTF-8.

There are no particular restrictions on the chicken to be immunized, but from the standpoint of mass production of antibodies, egg breeds such as white leghorn are preferably used.

In addition, immunization with GTF-3 can be carried out by conventional methods such as subcutaneous injection, intramuscular injection, intraperitoneal administration, etc., and methods such as nasal and eye drops. Furthermore, GTF-3
The dosage may be appropriately selected so as to obtain the desired antibody titer and not have any adverse effects on the chickens.

Usually, antibodies that specifically react with the administered antigen are obtained in chicken eggs (egg yolks) several weeks after the initial immunization.

Note that, if necessary, an adjuvant such as FCA (complete Freund's adjuvant) or FIA (incomplete Freund's adjuvant) may be used together with GFT-3.

The anti-GTF-3 antibody of the present invention can be prepared from eggs collected from chickens that have been immunized for at least one month.

In addition, the antibody titer in egg yolk was determined by enzyme-linked immunosorbent assay (ELISA).
, radioimmunoassay, etc., and the change in antibody titer can be tracked by measuring the antibody titer at intervals of about two weeks after immunization.

In the Examples described below, the anti-GTF-3 antibody of the present invention was prepared by tracking the change in antibody titer by measuring with EL[SA, and collecting eggs at a stage when the antibody titer had sufficiently increased.

Moreover, high antibody titers can usually be obtained for about 3 months.

In addition, if a decrease in the antibody titer is observed after immunization, the antibody titer can be increased by appropriately administering booster immunizations at appropriate intervals.

The high GTF-3 antibody of the present invention can be obtained, for example, by extracting and separating immunoglobulin contained in the egg yolk of a chicken immunized as described above. As this extraction/separation method, various commonly used methods for extracting/separating immunoglobulins can be used, such as a precipitation method using dextran sulfate or polyethylene glycol, or an extraction method using punopanol or chloroform. .

The anti-GTF-8 antibody of the present invention obtained as described above specifically reacts as an antibody with GTF-3 produced by 3.@utans. That is, it has enzyme activity inhibitory activity against GTF-8.

The anti-GTF-3 antibody of the present invention having an inhibitory activity against GTF-3 produced by S. mutans includes S. mutans.
By inhibiting the attachment of S+mutans to the tooth surface, the activity of S+mutans in the oral cavity is controlled,
It can prevent corrosion.

As described above, the present invention can be prepared more easily and in large quantities than the usual production method for preparing antibodies from mammals, and moreover, the RGC fraction of immunoglobulin can be prepared by an advanced operation of preparing antibodies from egg yolk. The object of the present invention is to provide an antibody that can be specifically isolated and purified, and which is useful as an active ingredient of an anti-erosive agent.

These various antibody-containing fractions can be mixed with a conventional dental caries preventive agent to prepare the dental caries preventive agent according to the present invention.

That is, the dental caries preventive agent of the present invention can be used only in oral products such as toothpastes such as toothpaste, powdered toothpaste, and liquid toothpaste, mouthwash, oral pasta, gingival pine surge cream, gargling tablets, troches, chewing gum, and canned drinks. not,
For that purpose, it is also applicable to various foods.

The amount of the antibody of the present invention to be added to the caries preventive material may be appropriately selected according to the dosage form thereof, and for example, 1
The amount of antibodies having an antibody titer of 0' or more can be about 0,0001 to 10% by weight.

As other components of the preventive agent of the present invention, appropriate components are used depending on the purpose of use, form of use, etc. For example, in the case of toothpaste, calcium carbonate, calcium hydrogen phosphate, calcium pyrophosphate, insoluble Sodium metaphosphate, hydrated alumina.

Abrasives such as silicic anhydride, glycerin, and sorbitol.

Moisturizers such as propylene glycol, sodium lauryl sulfate, and sodium lauroyl sarcosine.

It is produced according to a conventional method by mixing foamed rhododendrons such as stone powder, a binder such as sodium carboxymethyl cellulose, and carrageenan, and ingredients such as appropriate flavoring ingredients, sweeteners, and preservatives with water. Also, in mouthwashes and other mouthwashes, ingredients are appropriately blended depending on the properties of the product.

In addition, in the present invention, tooth stain removal agent 1 bad breath prevention agent,
It is also possible to incorporate various medicinal ingredients such as anti-caries agents such as fluorine and anti-enzyme preventive agents.

Therefore, the preventive agent of the present invention uses the egg yolk antibody prepared from the immunized eggs to inhibit Streptococcus.
Effectively inhibits plaque formation caused by mutans,
a! 4. Good prevention of eclipse. Furthermore, since the egg yolk antibody is highly safe, the preventive agent of the present invention is highly safe in use.

〔Example〕

The present invention will be explained in more detail with reference to Examples below.

In addition, the % display below indicates weight/volume % unless otherwise specified.

Example 1 Preparation of +al antigen S, 5utans Ingbritt strain (serotype C
1 (obtained from Osaka University School of Dentistry) was cultured in 13jl of BHIi3 analysis medium for 37t for 18 hours. The culture solution was separated into bacterial cells and culture supernatant by continuous centrifugation. The separated culture supernatant was subjected to ammonium sulfate precipitation treatment at 50% saturation, and the resulting precipitate was collected by centrifugation. Next, this precipitate was
A solution dissolved in istidine-HCI buffer (pH 6,2) was dialyzed against the same buffer, and the precipitate formed in the solution was removed by centrifugation, and the supernatant was diluted with polybuffer PBE94. (Polybuffer
PBE94, Pharmacia! Rino 2.5X25cm
column.

Both fractions adsorbed on the column were washed with Varibuffer 74 (Pol
ybuffer 74, manufactured by Pharmacia)
It was selectively eluted with a l+ gradient.

When the GTF activity of each eluted fraction was measured by the method described below, and the protein content was measured by 280rs + ultraviolet absorption method, significant GTF was found in both fractions of 9N5.4 to 5.0.
Activity was observed.

Measurement of GTF activity; sample 10μ! and 20sM (14C-glucose)s as a substrate.
ucrose) (0.05 ci/mol).
2M phosphoric acid solution (ρ) 16.0) Dull
After reacting at 37°C for 1 hour, the entire reaction solution was filtered through filter paper (
1, OX 2. After washing this with methanol, the amount of radioactivity incorporated into the methanol-insoluble glucan remaining on the filter paper was measured, the GTF activity was calculated, and 1 μm O1 of glucose was transferred to the glucan per minute. 1 unit (U) of activity.

Next, the elution fractions of I)) 15.4 to 5.0 were collected and subjected to ammonium sulfate precipitation treatment at 80% saturation to obtain a precipitate. This is dissolved in 1On+M phosphate buffer (pH 6,0) and dialyzed against the same solution. Furthermore, the precipitate formed in the solution was removed by centrifugation to obtain a supernatant. This supernatant liquid was used as a crudely purified specimen (immunization antigen).

The protein concentration and total protein amount of the sample were determined by CBB-
G method (Branford, M, M, + Anal, B
iochem,.

72, 248. (1976) ), the protein 1t tH degree was 0.4 mg/lel, and the total protein amount was 9.4 μg.

Furthermore, the specimen was subjected to 5DS-PAGE under the above-mentioned operating conditions.
When the sample was subjected to staining with Kumasi Brilliant Blue (CBB), several bands of low molecular weight, mainly a band at the 155 Dalton position, were detected.

Further, the GTF-3 content in the sample calculated from the above 5DS-PAGE results was 20% by weight.

Separately, activity staining was performed as described below. That is, the sample was treated in Tris-HCl buffer at 37°C for 30
After 5DS-PAGE in the same manner as described above, except that the electrophoresis was performed at 4°C, the gel was run at 37°C.
The soaked gel was soaked for 18 hours in a phosphate buffer (pH 6,0) containing 1% sucrose and 0.05% sodium azide at
When the sample was stained by odjcacid 5chiff) reaction, a prominent stained band indicating glucan production was detected at the 155 Dalton position.

Mountain) Immunization of antigen to chickens + crudely purified immune antigen preparation obtained by a+ 1. OmA(CBB-
Contains 0.4mg protein amount when measured by G method)
and FCA (Complete Freund's Adjuvant)], Omj
! were mixed 1:1 to form a W2O type emulsion.

Apply the obtained emulsilan to the breast muscle of the chicken.
After the first immunization, follow the method below.
Measure the antibody titer of WSF (described later) obtained from the collected eggs,
We observed its progress.

Next, as shown in FIG. 1, it was noticed that the antibody titer in the egg yolk started to decrease 8 weeks after the first immunization, and a second immunization was performed in the same manner as the previous time.

Eggs produced by the chickens were collected approximately one month after the completion of the secondary immunization.

(c) Preparation of antibodies Egg yolk isolated from eggs 13mj! and the same amount of PBS (
Phosphate buffer, pH 7,4) and add the same mixture to the resulting mixture! (26 m6) of chloroform was added and this was stirred well.

After stirring, the mixture was allowed to stand at room temperature for 30 minutes, and then centrifuged at 3.00 Orpm for 20 minutes to collect the top layer, the transparent fraction, and the antibody-containing fraction (Wate).
r 5olable Fracton: W S F )
And so.

These two parts are 2. It had an antibody titer of 1X10'.

In addition, WSF (13 blocks) showing the antibody titer of 2.1 x 10'
'; I 3vg1. The protein content of the WSF (prepared from the egg yolk of 3+aj!
).

td) Method for measuring antibody titer; The antibody titer was measured by EL[SA.

First, the purified immune antigen preparation obtained in Example 2 was added at 10 μg/r.
50mM sodium carbonate buffer (pH
The solution obtained by dissolving in 9,6) was placed in a 96-well plate [Immulon 2. 100 μE was added to each well of a tube (manufactured by Guinatec), left overnight at 4°C, and PBS (pH 7) containing the purified antigen contained in both aliquots was added.
, 4) at 37° C. for 1 hour to perform blocking, and then the plate was washed 5 times with PBS-T.

Here, 100 μl of the previously obtained two-step dilution of WSF in PBS-T was added to each well and reacted at 37° C. for 1 hour.

After the reaction, the plate was washed 5 times with PBS-T, and a peroxidase-conjugated anti-chicken IgG antibody (protein mass 1.67 μg/mj) was added to the plate as a secondary antibody.
1001IJ was added to each well, reacted at 25°C for 30 minutes, and then washed 5 times with PBS-T.

Next, add 50 ml of disodium phosphate-0.1 M citrate buffer (pH 15.0) to each well of the plate for 0.2 h.
mj! 100 μl of a solution containing 20 mg of the substrate O-phenyl diamine and 10 μl of hydrogen peroxide was added to the mixture, and the mixture was reacted at 25° C. for 20 minutes.

To stop the reaction, use 100μ of 3N sulfuric acid solution! I did it by adding.

After stopping the reaction, the antibody titer was measured by measuring the absorbance (ODl, t) of each well. The antibody titer was determined by the end point titer method, and the dilution ratio was set to give an absorbance of 0 or 2.

(f! Recognition of water-soluble glucan synthesis inhibitory activity of GTF-3 by el antibody) Using the antibody obtained with tc+, an experiment was conducted to inhibit the activity of GTF-3 produced by S, s+utans.

First, (W with an antibody titer of 2,1X10' obtained in C1)
SF and its 8-fold dilution were used as test solutions.

Separately, WSF was prepared from eggs of non-immunized chickens (the same chickens as in (b)) in the same manner as in 101 and used as a test solution. In addition, it was confirmed that the WsF did not specifically react with GTF-3.

In addition, phosphorus #Ill street solution (PBS) was used as a control.

Next, after dispensing 5 μl of each of these test solutions into the wells of a disposable replate (manufactured by Sanko Pure Chemical Industries),
Mo, 3*U of GT F-35II was added to each well.

After mixing, the mixture was allowed to react at 37° C. for 30 minutes.

After the reaction is complete, add 2011I to each well in the same manner as above.
Add 10μ of [14C-gluco,su]sucrose of M and react again at 37°C for 1 hour. The entire reaction solution was spotted on a filter paper and washed with methanol. The amount of radioactivity incorporated into the cells was measured, and GTF activity was calculated.

Table 1 shows the effects of each WSF, taking the GTF activity when PBS was added as 100%.

Example 2 Preparation of GTF-3 Purification Standard BHI of 151
Example 2 A supernatant solution corresponding to a crudely purified sample was obtained under the same conditions as in Example 1 except that a dialysis medium was used.

The supernatant was added to hydroxyapatite (Bio Gel) equilibrated with 1 (laM phosphate buffer (pH 6,0)).
IITP.

Column (1, manufactured by Bio-Rad)
0x13cm).

The percentage adsorbed on the column was selectively eluted by a concentration gradient of 0.01-0.56 phosphate buffer (pH 16,0).

The GTF activity and protein content of each portion were measured using the same method as described above.
GTF activity was observed in both cases.

Next, the eluted fractions showing high activity with 0.2M phosphate buffer (pH 6.0) were collected and dialyzed against 10+wM phosphate buffer (all 6.0) to prepare purified enzyme preparations. I got it.

Furthermore, when the sample was subjected to 5DS-PAGE and stained with Coomassie brilliant blue (CBB), a single band was detected at the Dalton position of 155.
The purified enzyme preparation was confirmed to be an enzyme protein with a molecular weight of 155 Daltons.

Furthermore, an amount of the purified enzyme preparation corresponding to 50+sU enzyme activity was added to the final concentration in 0.1H phosphate buffer containing 1% sucrose and 0.1% sodium azide (all 6
．． 0) and adjusted the volume to 3 ml with distilled water,
The mixture was mixed and the enzyme reaction was carried out at 37°C for 18 hours. In addition,
The enzyme reaction was stopped by cooling the reaction solution with water to 4°C.

After the enzymatic reaction was completed, the reaction product was centrifuged at 1600°g and separated into a precipitated water-insoluble fraction and a supernatant.

The water-insoluble fraction was washed twice with distilled water and suspended in 3 ma distilled water to obtain a water-insoluble glucan sample.

In addition, 2.5 times the volume of ethanol was added to the upper TN solution, and after leaving it at 4°C for 1 hour, the resulting precipitate was collected by centrifugation at 1,600g, dissolved in 3aIil of distilled water, and further The same precipitation treatment was performed again, and the recovered precipitate was dissolved again in 3 tal distilled water to obtain a water-soluble glucan sample.

Furthermore, as a result of quantitative analysis of the amount of glucan contained in these preparations using the Anthrone method, the amount of glucan contained in the water-insoluble glucan preparation was 0.05B/ml, and the amount of glucan contained in the water-soluble glucan preparation was 2.
．． A value of 6 mg/lll was obtained. When the type of glucan synthesized by the action of the purified enzyme preparation was investigated, it was revealed that the purified enzyme preparation was GTF-3, which has the ability to synthesize water-soluble glucan.

In addition, when the protein amount of the obtained purified enzyme preparation was measured by the above-mentioned method, it was 5.6B.

Furthermore, an antibody was prepared in the same manner as in Example 1 except that the purified GTF-3 immune antigen preparation was used as the antigen, and an antibody that specifically reacted with GTF-3 was obtained in the same manner.

Furthermore, as a result of testing the GTF-3 activity inhibiting effect of the obtained antibody using the same method, similarly excellent effects were observed.

Next, Examples of the dental caries preventive agent of the present invention will be described.

(Example 3) Toothpaste calcium pyrophosphate 42% glycerin 15% sorbitol 70%
10% carboxymethyl cellulose 1
2% Sankarin Sodium 0.1% Sodium Lauryl Sulfate 2.0% Fragrance
1.0%
The antibody-containing fraction (WSF) obtained in Example 1 was added to the component with a residual water content of 100% or more.
．． 5% (antibody titer 2.1xlO').

(Example 4) Moushwasosh ethanol 22.5% Sankarin sodium 0.05% Laurylgetanol 7mide 0.3% Fragrance
1.0% water
100 remaining
The fraction (WSF) containing the antibody obtained in Example 1 was 0% or more.
．． 5% (antibody titer 2.1 x 10').

(Effects of the Invention) The present invention has a sufficient inhibitory effect on the adhesion of S. mutans to the tooth surface, is excellent in mass production, has low production costs, is excellent in safety, and is a caries preventive agent. An antibody useful as an active ingredient and a caries preventive agent containing the antibody have been provided.

In particular, the antibody having immunological activity against S. mutans of the present invention has the following advantages compared to conventional antibodies obtained by immunizing mammals.

(1) The antibody of the present invention is obtained from the eggs of immunized chickens, and special methods are used for egg collection, egg handling, and antibody acquisition from rabbits.
Or it doesn't require any skill.

Furthermore, since only the IgG class of immunoglobulins is transferred to egg yolk, only IgG can be easily obtained.

On the other hand, when antibodies are obtained by blood collection from immunized mammals, skilled blood collection techniques are required, and it is extremely difficult to separate and purify large amounts of IgG from serum.

(2) The chickens used for preparing the antibodies of the present invention are easy to manage, and their management costs are low compared to, for example, rats.

Moreover, it is difficult to continuously obtain large amounts of blood and milk from mammals, and methods using mammals are not suitable for mass production of antibodies, but chickens can continue to produce eggs stably over a long period of time. Therefore, the antibody of the present invention can be mass-produced and the production cost is low.

(3) The stability of antibodies prepared from the blood or milk of immunized mammals is not necessarily good, and even in serum or in a purified state, the stability is not necessarily good, and even in serum or in a purified state, it is about -80°C It is necessary to store the product under the following temperature conditions.

The antibody of the present invention has good stability and good storage stability, and can be stored, for example, in egg form at 4°C for 1 to 2 months.

Finally, when preparing antibodies using chickens, it is not always possible to obtain a sufficient amount of antibodies that specifically react with the immunized antigen. For example, even when immunized with a viral antigen, a sufficient antibody titer could not be obtained.

Therefore, even if chickens are immunized with S. mutans bacterial surface protein antigen, it is completely unpredictable whether antibodies with a practical level of antibody titer will be obtained, and this was achieved for the first time by the present inventors. It is something.

[Brief explanation of the drawing]

FIG. 1 is a graph showing the change in antibody titer of immunized chickens in Example 1.

Claims (2)

[Claims] (1) The serotype of the pathogenic bacteria that causes caries is c, e, or
An immunoglobulin prepared from eggs produced by chickens immunized with water-soluble glucan synthase produced by Streptococcus mutans, which is f., and an antibody having inhibitory activity against the enzyme. (2) The serotype of the caries-inducing pathogen is c, e, or
An immunoglobulin prepared from eggs produced by chickens immunized with water-soluble glucan synthase produced by Streptococcus mutans, which is f. agent.