JPH01242534A - Antibody and anticarious agent containing said antibody as active ingredient - Google Patents

Abstract

PURPOSE:To obtain an antibody, consisting of immunoglobulin prepared from eggs produced by a hen immunized with a water-insoluble glucan synthetase produced by Stretococcus.mutans, capable of inhibiting sticking of the Streptococcus.mutans to tooth surfaces and useful as an active ingredient of an anticarious agent. CONSTITUTION:An antibody prepared by culturing Streptococcus.mutans having a serotype of a, d or g as a cariogenic pathogenic germ in a culture medium containing at least glucose to provide a water-insoluble glucan synthetase (TGF-1) from the culture supernatant thereof, immunizing a hen (preferably egg breed, such as White Leghorn) using the above-mentioned enzyme as an antigen, collecting eggs from the hen after the passage of >=1 months from the immunization and preparing the antibody having inhibitory activity against the TGF-1 therefrom. An IgG fraction can be specifically separated and purified by simple operation, such as migration of only the IgG fraction in immunoglobulin into egg yolk and preparation of the antibody from the egg yolk and a high antibody value can be normally obtained for about 3 months.

Description

[Detailed description of the invention]

3. Industrial application field The present invention is directed to the use of Streptococcus mutans (Streptococcus mutans), which is a pathogenic bacterium that induces U-erosion.
The present invention relates to an antibody having an inhibitory activity against water-insoluble glucan synthase (glucosyltransferase, abbreviated as GTF-1) produced by A. [Prior art] It is already known that the attachment process of S. mutans to the tooth surface plays an important role in the development of portal cavities, and the process of S. mutans attachment to the tooth surface plays an important role. Various attempts have been made to prevent portal erosion by controlling the adhesion. For example, a method for preventing crane erosion using an antibody having immunoactivity against S. somutans is known, and British Patent No. 1,505,513 discloses that breast milk obtained from a cow immunized with S. somutans is treated with S. somutans. IIutan
A method used to control the intraoral colonization of S.
Publication No. 38327 describes antiserum and/or milk obtained by immunizing mammals with cell walls obtained from culture and cultivation of S. A dental caries preventive agent is described which is a combination of one or more types of cilantrope selected from the group consisting of: [Problems to be Solved by the Invention] However, the effect of conventional antibodies having immunoactivity against S. mutans is not necessarily sufficient to prevent the development of bacterial infections. Since it is prepared from serum, it has drawbacks such as lack of mass productivity and high production cost, and is often impractical. The present inventors have conducted various studies to solve the problems in the prior art, and as a result, we have developed a method for preparing antibodies using chickens that has S. muta, which has the physicochemical properties described below.
By using water-insoluble glucan synthase of ns in combination as an antigen, S. The present invention was completed based on the new finding that it is possible to prepare an antibody that has a sufficient inhibitory effect on the adhesion of S. mutans to tooth surfaces and does not have the above-mentioned drawbacks. In particular, when preparing antibodies using chickens, it is not always possible to obtain enough antibodies that specifically react with the immunized antigen. For example, when immunizing with a virus antigen, a sufficient antibody titer may not be obtained. Therefore, when immunizing chickens, it is necessary to carefully examine the antigen and make sure to obtain one with a high antibody titer. The present inventors were the first to obtain the knowledge that antibodies with sufficiently high antibody titers can be obtained by immunizing chickens. , an antibody that has a sufficient inhibitory effect on the adhesion of S. mutans to the tooth surface, is excellent in mass production, has a low production cost, is excellent in safety, and is useful as an active ingredient of a preventive agent for portal caries, and contains the antibody. 1. An object of the present invention is to provide an anti-eclipse agent. [Means for solving the problem] The antibody of the present invention has inhibitory activity against water-insoluble glucan synthase (hereinafter abbreviated as GTF-1) produced by S. muLans. , S whose serotype is a, d or g having the following physicochemical characteristics, G produced by mutans
It can be obtained as immunoglobulin prepared from eggs produced by chickens immunized with TF-1. 0 action and substrate specificity It acts on sucrose and synthesizes water-insoluble glucan. (2) Molecular weight: The molecular weight measured by 5OS-polyacrylamide gel electrophoresis is 155 to 170 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 (
SDS-PAGE) was performed according to the following procedure. 5DS-PAGE operating conditions: 5DS-PAGE is performed by the method of Laemml et al. That is, each of the crude or purified samples described below was added to 2% S.
DS, 62.5 mM Tris-HCl buffer (pH 6) containing 5% 2-mercaptoethanol and 20% glycerol.
, 8) for 13 minutes at 100°C. Electrophoresis is performed in a 7.5% separation gel containing 0.1% SDS and a 4% concentration gel at room temperature at 1O2 for 2 hours. In addition, as molecular weight markers, ferritin (ferri-mouth n, 220 Dalton), phosphorylase b (phosphory-1ase b, 9
Dalton 4), bovine serum albumin (bovinese
rum albumin, 67 k Daltons), catalase (60 k Daltons),
ovalbumin (43 kdalton) and lactate dehydrogenase (lactate dehydrogenase)
Drogenase. Coomassie Brilliant Blue (CBB) was used for band detection.
) can be carried out by staining with. As GTF-1 used to obtain the antibody of the present invention (hereinafter referred to as anti-GTF-I), any enzyme having the above characteristics can be used. It can be obtained by purifying it from the liquid. That is, Slmutans having serotypes a, d, or g are cultured in an appropriate medium and purified from the culture supernatant. The G-type seedlings used here include S, mutans 67
15 strain, ATCC27351 strain, etc., which are known and easily available, can be used, for example, S, muta
ns 6715 strain was obtained from Osaka University School of Dentistry and ATCC27.
351 stocks are American Type Cultures Co.
11ection (ATCC)). Also, for type A and type D seedlings, known strains may be similarly obtained and used. Further, as a medium, a medium containing at least glucose can be used, for example, TTY mound (Trypticase).
, TryptoseSYeast extract
complex medium), BHI (Brain Hear
Infusion medium, FMC medium, etc. can be used. In addition, the culture temperature may be within a range that allows bacterial cell growth and is suitable for the production of GTF-[, but from the viewpoint of good bacterial cell growth and GTF-I production, it is usually 37°C. It is best to set it as a degree. 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-1 is reached, and may be about 18 to 24 hours. Further, other culture conditions may be appropriately selected from the above viewpoint. Next, GTF-1 is extracted from the bacterial cells. GTF-1 of S. mutans is Furuta+T
, et al.'s method (Furuta, T, +et al;
J, General old crobiology. , 131, 285-293 (198!i)) 4. :
Therefore, BHI dialysis medium was inoculated with S. 5utans4,
After growth, the bacterial cells were removed by centrifugation, and ammonium sulfate was added to the supernatant to precipitate the 50% ammonium sulfate fraction.
After dialyzing against istidine-11cI and performing chromatofocusing column chromatography, a purified sample can be obtained by performing hydroxyapatite column chromatography. As the antigen for obtaining antibodies, the bacterial cell culture supernatant or ammonium sulfate 1 reduced sample obtained by the above operations, the crudely purified sample after chromatofocusing, or the purified sample G
TF-1 etc. can be used. Next, the method for preparing the anti-GTF-1 antibody of the present invention will be described in detail. The anti-GTF-1 antibody of the present invention can be prepared from eggs of chickens immunized with the above-mentioned GTF-■. 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. Immunization methods with GTF-1 include conventional methods such as subcutaneous injection, intramuscular injection, and intraperitoneal administration, as well as methods such as nasal and eye drops. I can do it. Furthermore, GTF
The dose of -1 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.
' If necessary, an adjuvant such as FCA (Freund's complete adjuvant) or FIA (Freund's incomplete adjuvant) may be used together with C; FT-1. The anti-GTF-1 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 measured using enzyme-linked immunosorbent assay (ELIS^).
, 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-I antibody of the present invention was prepared by tracking the change in antibody titer by measuring with ELISA, 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 small amount of antibody titer is found after immunization, the antibody titer can be increased by appropriately administering booster immunizations at appropriate intervals. The anti-GTF-T 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. Examples of this extraction/separation method include precipitation methods using dextran sulfate or polyethylene glycol, extraction methods using propatool or chloroform, and various other commonly used methods that can extract and separate immunoglobulins. can. Anti-GTF- of the present invention obtained as described above

[The antibody specifically reacts as an antibody against GTF-1 produced by S. mutans. That is, it has enzyme activity inhibitory activity against GTF-T. The anti-GTF-I antibody of the present invention having inhibitory activity against GTF-1 produced by S. mutans is
By inhibiting the attachment of tans to the tooth surface, S,
The intraoral activities of mutans can be controlled and crane erosion can be prevented. As described above, the present invention enables preparation of antibodies in large quantities more easily than in conventional production methods from mammals, and moreover, only the IgG fraction of immunoglobulin can be prepared by the simple operation of preparing antibodies from egg yolk. The object of the present invention is to provide an antibody that can be specifically separated and purified, and is useful as an active ingredient of a preventive agent for dental caries. These antibody-containing fractions can be mixed with a conventional AI corrosion preventive agent to prepare the anti-corrosion agent according to the present invention. That is, the M anti-caries agent of the present invention can be used in the oral cavity, such as toothpaste, toothpaste, powdered toothpaste, liquid toothpaste, etc., mouthwash, oral pasta, gingival nosage cream, gargling tablets, troches, chewing gum, canned drinks, etc. Not only products,
For that purpose, it is also applicable to various foods. The amount of the antibody of the present invention to be added to the crane erosion preventive material may be appropriately selected according to the dosage form, and for example, 1
0 for antibodies with an antibody titer of 03 or higher, OOO1 to 10
It can be about % by weight. As other components of the dental caries prevention agent of the present invention, appropriate components can be 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, and hydrated alumina. Abrasives such as silicic anhydride, glycerin, and sorbitol. Moisturizers such as propylene glycol, sodium lauryl sulfate, and sodium lauroyl sarcosine. A foaming agent such as soap powder, a binder such as sodium carboxymethyl cellulose and carrageenan, and other components such as appropriate fragrance ingredients, sweeteners, preservatives, etc. are mixed with water and produced according to a conventional method. 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 dental caries prevention 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,
Good prevention of candy corrosion. Moreover, since the egg yolk antibody is highly safe, the caries 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 %, especially when it is not hygienic. Example 1 (a) Preparation of antigen S 9mutans 6715 strain (serotype g, obtained from Osaka University School of Dentistry) was incubated in 151 BIT dialysis medium at 37°C.
The cells were cultured for 118 hours. The culture solution was separated into bacterial cells and culture supernatant by continuous centrifugation. 50 for 1n on isolated culture
% saturation with ammonium sulfate precipitation, and the resulting precipitate was collected by centrifugation. Next, this precipitate was
A solution dissolved in dine-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 poured into polybuffer y- PBE94 (+'olybufferP
BE94, manufactured by Pharmacia) 2.5X25cmO
applied to the column. Both fractions adsorbed on the column were treated with Parihampha-74 (Pol
ybuffer 74, manufactured by Pharmacia)
l+gradient 1 then 0.1 M'a degree of Nacl? :Selectively eluted with i7 solution. When the GTF activity of each eluted fraction was measured using the method described below, and the protein content was measured using a 280 nm ultraviolet absorption method, it was found that there was significant GTF activity in both fractions of 0.1 M Nac I mW. Admitted. Measurement of GTF activity; 10 μn of sample was added to 20 mM [14C-glucose]sucrose ([14C-glucose]s) as a substrate.
ucrose) (0,05ci/No1) and 20
, mixed with 10 μf of 0.2 phosphate buffer (pl+ 6.0) containing 17 M dextran TIO and incubated at 37°
After reacting for 1 hour at C, the entire reaction solution was filtered through filter paper (+,
．． After washing this with methanol, the amount of radioactivity incorporated into the methanol-insoluble glucan remaining on the filter paper was measured, and the GTF activity was calculated.
The activity of transferring 1 μmol of glucose to glucan per minute was defined as 1 unit (U). Next, the elution fractions of 0.1 M Nac at 14 degrees were collected and subjected to ammonium sulfate precipitation treatment at 80% saturation to obtain a precipitate. Add this to 10mM phosphate buffer (pf+6゜0)
and dialyze against the same solution. Furthermore, the precipitate formed in the mixture was removed by centrifugation to obtain a supernatant. This supernatant liquid was used as a crudely purified specimen (immunization antigen). 2K Protein concentration of 45 products and total protein 1C
BB-II; Law (Branford, M.M., An
al. Biochem. 72, 248. (1976)], the protein concentration was 0.4 mg/ml, and the amount of whole star was 13.2 g. Furthermore, the specimen was subjected to 5DS-r'AC under the aforementioned 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 160 Dalton position, were detected. Further, the GTF-1 content in the sample calculated from the above 5DS-PAGE results was 4.0% by weight. Separately, activity staining was performed as described below. That is, the preparation was treated in Tris-HCl buffer at 37°C.
After 5DS-PAGE with the same procedure as described above except that the electrophoresis was performed at 0 min and the electrophoresis was performed at 4°C, the gel was incubated with 1% sucrose and 0.05% dextran TIO and 0.05% dextran at 37°C. % sodium azide in phosphate buffer (pH 6,0) for 18 hours.
The immersion-treated gel is PAS (Periodic
When the sample was stained by Acid 5chiff) reaction, a prominent stained band indicating glucan production was detected at the 160 Dalton position. (b) Immunization of chicken with antigen I■ Purified immune antigen preparation obtained in (a) 1. Omj2 (CB
Contains 0.4 mg protein amount when measured by B-C method) and FCA (Freund's complete adjuvant) 1.0 m
j2 was mixed in a 1:l ratio to form a W2O type emulsion. After injecting 1.0 ml of the obtained emulsion into the chicken's pectoral muscle for initial immunization, measure the antibody titer of WSF (described later) obtained from the collected eggs according to the method below, and observe its progress. did. Next, as shown in FIG. 1, it was confirmed that the antibody titer in the egg yolk began to decrease 8 weeks after the first immunization, and a second immunization was performed in the same manner as the previous time. Approximately one month after the completion of the secondary immunization, eggs produced by the chickens were collected. (C) Antibody preparation Mix 13 ml of egg yolk separated from the egg with the same amount of PBS (phosphate buffer, al17.4), and add 1 ml of the same amount (26 mj2) to the resulting mixture. ) of chloroform was added and the mixture was diluted with <JAt↑. After stirring, the mixture was left at room temperature for 30 minutes, centrifuged at 3.00 Orpm for 20 minutes, the top transparent fraction was collected, and the antibody-containing fraction (Hate
r 5olable FracLon: W S F )
And so. Both portions had antibody titers of 0.7 x 10'. In addition, WSF (1
The protein concentration of 3ml (prepared from 13mf of egg yolk) was measured by Piuret's method, and the total protein mass in the WSF was determined to be approximately 26mg (approximately 2.0mg/mj2 13ml). (d) Method for measuring antibody titer; Antibody titer was measured by ELISA. First, 2.5 μg/g of the purified immune antigen preparation obtained in Example 2 was prepared.
50mM sodium carbonate buffer (
pl! The solution obtained by dissolving in 9.6) was added to a 96-well plate [Immulon 2 + Guinatenku! ! ! ] Add 100 μl to each well of
The plate was left at 4'C overnight to allow the purified antigen contained in the cough fraction to be adsorbed onto the plate. The plate was then washed five times with PBS-T (PBS containing 0.05% Tween 20, PH 7.4). After washing, the plate was washed with PB containing 3% BSA (bovine serum albumin).
After blocking by contacting with S (pH 7, 4) at 37°C for 1 hour, the plate was washed 5 times with PBS-T. Here, 100 μP of the previously obtained two-step dilution of WSF in PBS-T was added to each well, and the reaction was carried out at 37° C. for 1 hour. After the reaction, the plate was washed 5 times with Pr3S-T, and the plate was further injected with verokindase-conjugated anti-chicken IgG antibody (protein ffi+, 67t) as a secondary antibody.
Add 100 μN of tg/mff1) to each well and
After reacting at 5°C for 30 minutes, it was washed 5 times with PBS-T. Next, add 0.2 portions of disodium phosphate-0.1M citrate buffer (pl+5.0) to each well of the plate.
100 μP of a solution containing 20 mg of the substrate O-phenylenediamine and 10 μl of hydrogen peroxide was added to 0 m 2 and reacted at 25° C. for 20 minutes. The reaction was stopped by adding 100 μl of 3N sulfuric acid solution. After stopping the reaction, the antibody titer was measured by measuring the absorbance (00492) of each well. The antibody titer was determined by the end point tie method, and the dilution ratio was set to give an absorbance of 0.2. (e) Confirmation of the activity of GTF-1 to inhibit water-insoluble glucan synthesis using the antibody (C) Using the antibody obtained in (C), an experiment was conducted to inhibit the activity of GTF-1 produced by S. mutans. First, WSF having an antibody titer of 0.7 x 10' obtained in (C) and its 8-fold dilution were used as test solutions. Apart from this, non-immunized chickens (mountains) and similar chickens)
WSF was prepared from the eggs in the same manner as in (C) and used as a test chicken. In addition, it was confirmed that 1.5 WSF does not specifically react with GTF-I. In addition, phosphate buffered saline (PBS) was used as a control. Next, after dispensing 250 μl of each of these test solutions into 13 x 100 test tubes, further add 0.3n+Ll of GTF.
-T250μ2 was added to each tube. After mixing, the mixture was allowed to react for 30 minutes at 37°C. After the reaction is complete, add 2% sucrose and 40% sucrose to each test tube.
After adding 5007al of 0.2M phosphate buffer containing μM dextran TIO and reacting again at 371C for 1 hour,
The total amount of the reaction solution was ice-cooled to 4@C, subjected to ultrasonication, the turbidity (ODss.) of the reaction solution was measured, and the water-insoluble glucan synthesis 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 purified GTF-1 source product Under the same conditions as in Example 1, a supernatant liquid corresponding to a crudely purified sample was obtained. The supernatant was equilibrated with 11011I'J acid buffer (pl+6.0) to prepare hydroxyapatite (Bio Gel).
IITP, manufactured by Bio-Rad]
column (+, OX13cm). Both fractions adsorbed on the column were selectively eluted using a concentration gradient of 0.01 to 0.5 Ml77 acid buffer (pH 6, o). The GTF activity and protein content of both samples were measured using the same method as described above.
GTF activity was observed in both the samples (6, 0). Next, the eluted fractions showing high activity with 0.2 phosphate buffer (pl + 6.0) were collected and dialyzed against 10 mF phosphate buffer (pH 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 160 Dalton position.
It was confirmed that the purified enzyme preparation was an enzyme protein with a molecular weight of 160 Daltons. Furthermore, an amount of the purified enzyme preparation corresponding to 50 m[+ enzyme activity was added to a final concentration of 0.1 M phosphate buffer containing 1% sucrose and 0.1% sodium azide (ρI+6.
After adjusting the volume to 3 ml with distilled water, the mixture was mixed, and the enzymatic reaction was carried out at 37°C for 118 hours. In addition,
The enzyme reaction was stopped by cooling the reaction solution on ice to 4°C. After completion of the fermentation reaction, the reaction product was centrifuged at 1600xg and separated into the precipitated water-insoluble fraction and the supernatant. The water-insoluble fraction was washed twice with distilled water and suspended in 3 mE distilled water to obtain a water-soluble glucan sample. In addition, 2.5 times the volume of ethanol was added to the supernatant,
After standing at 4°C for 1 hour, the resulting precipitate was collected by centrifugation at 1600 g, dissolved in 3 ml of distilled water,
Furthermore, the same precipitation treatment was performed again, and the recovered precipitate was dissolved again in 3 ml of distilled water to obtain a water-soluble glucan sample. Furthermore, we quantitatively analyzed the amount of glucan contained in these preparations using the Anthrone method to investigate the types of glucans synthesized by the action of the purified enzyme preparations, and found that the purified enzyme preparations synthesized water-insoluble glucans. GTF has the effect of
□It became clear that it was I. Furthermore, when the protein amount of the obtained purified enzyme preparation was measured by the above-mentioned method, it was found to be 0.48 mg. When antibodies were prepared in the same manner as above, antibodies that specifically reacted with GTF-1 were obtained.Furthermore, the GTF-3 activity inhibiting effect of the obtained antibodies was tested in the same manner. As a result, similar excellent effects were observed. Next, an example of the 'gM C4 preventive agent of the present invention will be described. (Example 3) Toothpaste calcium pyrophosphate 42% glycerin 15% sorbitol 70%
10% carboxymethyl cellulose 12
% Sodium saccharin 0.1% Sodium lauryl sulfate 2.0% Flavoring
1. 0%
water
The antibody-containing fraction (WSF) obtained in Example 1 was added to the components with a remaining amount of 100% or more.
．． 5% (antibody titer 0.7 X IO') is blended. (Example 4) Moushuwonshu ethanol 22°5% Sankarin sodium 0.05% lauryl jetanolamide 0.3% fragrance
], O% water
Remaining ■
100% or more of the antibody-containing fraction (WSF
) 0.5% (antibody titer 0.I x 10')
Blend. (Effects of the Invention) The present invention has a sufficient inhibitory effect on the adhesion of S and mutans to tooth surfaces, is excellent in mass production, has low production cost, is excellent in safety, and is a caries preventive agent. An antibody useful as an active ingredient and a caries prevention agent containing the antibody have been provided. In particular, the antibody having immunological activity against S. muLans 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 collecting eggs, handling rabbits, and obtaining antibodies from rabbits.
Or a) It does not require sophisticated techniques. 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, horses, horses, etc. Moreover, 8 tons from l milk! It is difficult to continuously obtain large amounts of blood or milk, and methods using mammals are not suitable for mass production of antibodies. However, chickens continue to produce eggs stably over a long period of time, so the antibodies of the present invention can be used. is possible and the production cost is low. (3) Antibodies prepared from the blood or milk of immunized mammals do not necessarily have good stability, and even in serum or purified form, they must be stored at temperatures around -80°C. Ru. 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.

[Brief explanation of the drawing]

Figure 1 shows the antibody titer t of immunized chickens in Example 1.
It is a graph showing Ik shift. Procedural amendment to the body (method) July 1988 Patent Application No. 70331 2, Title of the invention 3, Relationship with the case of the person making the amendment Patent applicant address Tokyo 5-17-4 Sumida, Sumida-ku, Miyako 534
Kanebo Co., Ltd. 1-5-90 Tomobuchi-cho, Bejima-ku, Osaka City
Patent Department Telephone (06) 921-1251 6. Field of industrial application described in column 7 of "Detailed Description of the Invention" of the specification to be amended, content of the amendment 8tHt, page 1, line 18 ” shall be corrected as shown in the attached sheet. 8. List of attached documents (1) Attachment 1 Mail order Attachment 3, Detailed description of the invention [Field of industrial application]

Claims (2)

[Claims] (1)■ The serotype of the caries-inducing pathogen is a, d, or
An immunoglobulin prepared from eggs produced by chickens immunized with water-insoluble glucan synthase produced by Streptococcus mutans, which is Streptococcus mutans, and which has inhibitory activity against the enzyme. (2)■ The serotype of the caries-inducing pathogen is a, d, or
An immunoglobulin prepared from eggs produced by chickens immunized with the water-insoluble glucan synthase produced by Streptococcus mutans, which is Streptococcus mutans, and which contains as an active ingredient an antibody that has inhibitory activity against the enzyme. Preventive agent.