CN105418758A - Preparation method and application of anti-periodontosis antibody - Google Patents
Preparation method and application of anti-periodontosis antibody Download PDFInfo
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- C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
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- A23G1/00—Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor
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- A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
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- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
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Abstract
The invention provides a preparation method of an anti-periodontosis antibody, which comprises the following steps: taking porphyromonas gingivalis to prepare a thallus adjuvant antigen; mixing the porphyromonas gingivalis with the same amount of Freund adjuvant to prepare a thallus adjuvant antigen; taking 1-5ml of porphyromonas gingivalis antigen every time to carry out multi-point injection on mammals, and collecting serum to prepare a crude extract of an anti-porphyromonas gingivalis antibody IgG; and finally, carrying out centrifugal purification on the crude extract of the anti-porphyromonas gingivalis antibody IgG to prepare a pure product of the anti-porphyromonas gingivalis antibody IgG. According to the invention, the production cost of the antibody is reduced, and the titer of the antibody is increased. The invention also provides application of the anti-porphyromonas gingivalis antibody IgG. The anti-porphyromonas gingivalis antibody IgG prepared by the preparation method provided by the invention and antibacterial agents are used as effective components of passive immunization for preventing occurrence of periodontosis.
Description
Technical field
The invention belongs to medicine technology field, specifically a kind of preparation method of periodontopathy antibody.
Background technology
For oral disease, people expect that maximum is exactly carious tooth.But up-to-date investigation display, at present in developing country, the sickness rate of periodontopathy has exceeded the sickness rate of carious tooth, up to more than 75%.And the pathogenic bacterium that periodontopathy is main, be acknowledged as porphyromonas gingivalis.At present that porphyromonas gingivalis tropina that is single or mixing is made antigen, and the mammals such as immune horse cattle and sheep rabbit, extract antibody, carry out passive immunization control from animal serum to the measure that porphyromonas gingivalis carries out passive immunization.
Summary of the invention
The object of this invention is to provide the preparation method of a kind of anti-periodontopathy bacteria antibody IgG, the production cost of antibody is reduced, antibody titer improves.Another object of the present invention is the application of resisting porphyromonas gingivalis IgG antibody.There is provided the resisting porphyromonas gingivalis IgG antibody prepared using the present invention and antiseptic-germicide as the generation of passive immunization effective ingredient for prevention of periodontal disease.
The technical scheme adopted is:
1, a preparation method for anti-periodontopathy bacteria antibody, is characterized in that comprising the steps:
(1) porphyromonas gingivalis is got, Anaerobic culturel 5-7 days in BHI substratum; Cultured porphyromonas gingivalis is carried out centrifugal treating, collects thalline, wash thalline 4-6 time with the phosphate buffer normal saline that pH6-7, concentration are 0.05-0.2M, at the temperature of 50-65 DEG C, heating 25-35 minute, carries out inactivation treatment, obtains porphyromonas gingivalis liquid;
The freund adjuvant of porphyromonas gingivalis liquid and equivalent is mixed, uses high speed homogenizer process, obtained thalline adjuvant antigen;
(2), according to dried meat breast class animal species, get porphyromonas gingivalis antigen 1-5ml at every turn, lymphoglandula and dorsal sc multi-point injection are carried out to animal, inject three times altogether, every minor tick three weeks; After final immunization 20 days, to tire after testing qualified rear collection serum, get under immune serum stirs and add the dilution of appropriate salt solution, adjust ph, to 7.5-8.5, adds equivalent saturated ammonium sulphate solution, at 2-8 DEG C of temperature, leave standstill 20-30 hour, high speed centrifugation 20-30 minute, getting its precipitation, through melting ultrafiltration and concentration desalination, degerming postlyophilization again, obtaining resisting porphyromonas gingivalis IgG antibody crude extract;
(3), by after resisting porphyromonas gingivalis IgG antibody crude extract centrifugal purification, then through DEAE-SephadexA50 column chromatography; Carry out continuously or fraction gradient wash-out with the phosphoric acid buffer containing NaCl, collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, Function protein content is 20mg/ml;
By the protein eluate of previous step gained again through SephadexG200 gel filtration chromatography purifying, carry out wash-out with the phosphoric acid buffer containing NaCl, collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, degerming through 0.22 μ membrane filtration, lyophilize, obtains resisting porphyromonas gingivalis IgG antibody sterling.
The speed of above-mentioned centrifugal purification is 5000-12500rpm, centrifugal number of times 2-5 time, in DEAE anion exchange chromatography, the concentration of elutriant phosphoric acid buffer is 0.003-0.02M, the concentration of contained NaCl is 0.03-0.1M, gel filtration chromatography elutriant phosphate buffer density is 0.01-0.2M, and the concentration of contained NaCl is 0.1-0.5M; Affinity chromatography eluate concentration is 0.1-0.5M.
Above-mentioned porphyromonas is unicellular, uses BHI substratum, and under 37 DEG C of temperature anaerobic conditions, quiescent culture porphyromonas gingivalis 5-7 days, by centrifugal 10 minutes of porphyromonas gingivalis 4000rpm after cultivating, collects thalline; With pH value be 6.0, concentration is that the phosphate buffer normal saline of 0.2M washes thalline 5 times, heat inactivation process in 50 DEG C, 25 minutes, obtained porphyromonas gingivalis liquid.Get bacterium liquid (2,000,000,000/ml), mix with the freund adjuvant of equivalent, use high speed homogenizer process, obtained porphyromonas gingivalis adjuvant antigen.
According to mammal species, get thalline adjuvant antigen 1-5ml at every turn, lymphoglandula and subcutaneous multi-point injection are carried out to animal, inject three times altogether, every minor tick three weeks;
Third time immune animal after 20 days, detection tire qualified after, collect antiserum(antisera) salt solution to dilute, adjust ph to 8.0, add under stirring equivalent saturated ammonium sulphate solution put 2-8 DEG C leave standstill 24 hours, centrifugal 20 minutes of 8000rpm, gets precipitation, through ultrafiltration desalination, concentrated, degerming postlyophilization, obtained anti-periodontopathy bacteria antibody IgG crude extract;
By anti-periodontopathy bacteria antibody IgG crude extract through DEAE-SephadexA50 chromatography column chromatography; Post bed 2.5 × 35cm, adds the sample 3.0ml of 10mg/ml, carries out wash-out, flow velocity 20ml/h with 0.01M, pH7.0 phosphoric acid buffer containing 0.07MNaCl, and often pipe 5.0ml pressed by elutriant; Collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, Function protein content is 20mg/ml; Then by this liquid further through SephadexG200 gel filtration chromatography; Post bed 2.0 × 65cm, application of sample 1.5ml, carry out wash-out, flow velocity 8.0ml/h with 0.01M, pH7.0 phosphoric acid buffer containing 0.1MNaCl, and often pipe 5.0ml pressed by elutriant, collects each albumen elution peak, and detects the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, degerming through 0.22 μ membrane filtration, lyophilize, obtains resisting porphyromonas gingivalis IgG antibody sterling.
The application of the anti-periodontopathy antibody obtained by the inventive method:
Resisting porphyromonas gingivalis IgG antibody prepared by the present invention is for the additive of anti-periodontal germ product, and usage quantity is 5-20g/Kg.Its product is with the composition of the anti-periodontopathy bacteria antibody IgG anti-periodontopathy that is effective ingredient.
Porphyromonas gingivalis is the main bacteria seed causing periodontopathy, and has certain cross-immune reaction to other bacterial classification.Select porphyromonas gingivalis as major antigen thalline, avoid mixed bacterium when making antigen, the problem that Main Bacteria effect reduces, it also avoid single culture when making antigen, the problem that scope is partially narrow.Ensure that antibody has cross reaction in a big way.
Show that IgG antibody of the present invention reaches PAGE purity after testing, through molecular weight determination, IgG antibody molecular weight is 160,000 dalton.
IgG antibody physical and chemical experiment result shows, IgG antibody of the present invention, under 65 DEG C of conditions, still maintains activity in 90 minutes; In the environment of pH4-11, within 37 DEG C, 8 hours, its antibody activity is without considerable change, but antibody activity declines rapidly when pH2 and pH12, and very fast inactivation; IgG antibody of the present invention has larger impermeabilisation pressure character, can tolerate the sucrose osmotic pressure of 40%.
IgG antibody indirect hemagglutination Inhibition test shows, IgG antibody of the present invention can suppress porphyromonas gingivalis to condense effectively, and it tires up to 1:512; IgG antibody suppresses the observation of porphyromonas gingivalis adhesion experiment to show, IgG antibody of the present invention is diluted to 1:8, still has and significantly suppresses porphyromonas gingivalis at the adhesive attraction of Tooth surface; Experimentation on animals shows, feeds the big white mouse infecting porphyromonas gingivalis, effectively can prevent the generation of periodontopathy by IgG antibody of the present invention, detects and shows that bacterial plaque Porphyromonas gingivalis number have dropped 70-80%.
When anti-periodontopathy composition of the present invention is liquid state, spraying in small-sized portable type spray tank can be loaded and used.
IgG antibody of the present invention also can coordinate with preventives that decays tooth such as tooth staining removers, in advance anti-halitosis dose, and makes various control product.Also can join in various oral cleansing lotion, increase the anti-periodontopathy function of oral cleansing lotion.
Further embody rule comprises:
1, by resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, to add in potassium sorbate, Sodium Benzoate two kinds of antiseptic-germicides at least one as the anti-periodontopathy composition of effective ingredient, comprise toothpaste, suck liquid, the oral product of collutory and aerosol, or chewing gum, chocolate, ice cream, ice cream food, the add-on of IgG antibody of the present invention is 0.05-0.2%, and the add-on of potassium sorbate, Sodium Benzoate is 0.01-0.02%.
2, the resisting porphyromonas gingivalis IgG antibody adopting the inventive method to prepare, the method for liquid is sucked in preparation, is get IgG antibody 2.0g of the present invention, potassium sorbate 0.15g, Sodium Benzoate 0.15g, aspartame 0.8g, mentha camphor 0.15g, apple flavour 0.4ml; Mentha camphor added in the distilled water of 100ml, 60 DEG C dissolve; Other solid content is respectively dissolved in the distilled water of 450ml, is then mixed by the uniform liquid of various dissolving, with distilled water constant volume to 1000ml, to obtain final product.
3, by resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, prepare the method for chewing gum, get IgG antibody 2.0g of the present invention, potassium sorbate 0.15g, aspartame 0.8g, mentha camphor 0.15g, microencapsulation apple flavour 0.4g, matrix 10.0g, Walocel MT 20.000PV 5.0g, by total amount 1 kilogram, to obtain final product.
4, the resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, the method of toothpaste processed, get IgG antibody 0.1g of the present invention, potassium sorbate 0.015g, Sodium Benzoate 0.015g, glycerine 10.0g, Sorbitol Powder 8.0g, Walocel MT 20.000PV 2.0g, Thallus Gracilariae 1.3g, Sodium Lauryl Sulphate BP/USP 1.8g mentha camphor 0.015g, aspartame 0.015g, strawberry flavour 0.05ml, calcium phosphate two water thing 47.8g, by Walocel MT 20.000PV, Thallus Gracilariae adds in the distilled water of about 60ml, swelling is dissolved, successively add other composition again, abundant stirring, make to mix, finally add distilled water and be settled to 100ml, be stirred into paste, obtain.
5, by resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, the method for tooth cream is protected in preparation, is get IgG antibody 0.1g of the present invention, Sodium Benzoate 0.01g, beeswax 8.0g, stearic acid 10.0g Zerol 2.0g, glycerine 10.0g, Walocel MT 20.000PV 1.0g, mentha camphor 0.01g, aspartame 0.05g, distilled water 68.80ml, strawberry flavour 0.02g, by beeswax, stearic acid, Zerol, glycerine mixing, be heated to 70 DEG C, the liquid of mixed dissolution is referred to as A liquid; Walocel MT 20.000PV is dissolved in the distilled water of 50ml, swelling is dissolved, successively add IgG antibody, mentha camphor, aspartame, potassium sorbate, strawberry flavour again, abundant stirring, mix, then add cooled A liquid, finally add distilled water and be settled to 100ml, be stirred into paste, obtain final product.
IgG antibody technology of preparing production cost of the present invention is low, and antibody titer is high, impermeabilisation pressure, has stronger immunocompetence to porphyromonas gingivalis, and has a wide range of cross reaction to the pathogenic bacterium beyond porphyromonas gingivalis.Anti-periodontopathy composition of the present invention, has IgG antibody consumption little, use safety, effectively can prevent the generation of periodontopathy.
Embodiment
Embodiment one
Use BHI substratum, under 37 DEG C of anaerobic conditions, quiescent culture porphyromonas gingivalis 5-7 days, by centrifugal 10 minutes of porphyromonas gingivalis 4000rpm after cultivating, collection thalline pH6. concentration is that the phosphate buffered saline buffer of 0.2M washs thalline 5 times, through 25 minutes heating (50 DEG C) inactivation treatment, obtain porphyromonas gingivalis liquid.Get the freund adjuvant that bacterium liquid (2,000,000,000/ml) adds equivalent, use high speed homogenizer process, obtained water-in-oil shape thalline adjuvant antigen.
According to the different sorts of dried meat breast class animal, get thalline adjuvant antigen 1-5ml at every turn, lymphoglandula and the immunity of subcutaneous multiple spot are carried out to animal, altogether immunity three times, every minor tick three weeks.
After immune animal, blood sampling in 20 days measures antibody titer for the third time, start to collect antiserum(antisera) after reaching standard, dilute with salt solution, adjust ph to 8.0, adds equivalent saturated ammonium sulphate solution under stirring, put 2-8 DEG C and leave standstill 24 hours, centrifugal 20 minutes of 8000rpm, get precipitation, through ultrafiltration desalination, concentrated, degerming postlyophilization, obtained resisting porphyromonas gingivalis IgG antibody crude extract.
By resisting porphyromonas gingivalis IgG antibody crude extract through DEAE-SephadexA50 column chromatography.Column volume is 2.5 × 35cm, application of sample 10-50ml(10mg/ml), carry out wash-out with 0.01M, pH7.0 phosphoric acid buffer containing 0.07MNaCl, collect elutriant often pipe 5.0ml; Collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, Function protein content is 20mg/ml.Then by this liquid further through Sephadex gel filtration chromatography purifying.Column volume 2.0 × 65cm, application of sample 15-30ml, carry out wash-out with 0.01M, pH7.0 phosphoric acid buffer containing 0.1MNaCl, collects elutriant often pipe 5.0ml, collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA.Collect the protein eluate having antibody activity, degerming through 0.22 μ membrane filtration, lyophilize, obtains resisting porphyromonas gingivalis IgG antibody sterling.
The preparation of anti-periodontopathy composition product.
Embodiment two
IgG antibody sucks the preparation of liquid.Get IgG antibody 2.0g of the present invention, potassium sorbate 0.15g, Sodium Benzoate 0.15g, aspartame 0.8g, mentha camphor 0.15g, apple flavour 0.4ml.Mentha camphor added in the distilled water of 100ml, 60 DEG C dissolve; Other solid content is respectively dissolved in the distilled water of 450ml, then mixes, the uniform liquid of various dissolving with distilled water constant volume to 1000ml.
Embodiment three
The preparation of IgG antibody chewing gum.Get IgG antibody 2.0g of the present invention, potassium sorbate 0.15g, aspartame 0.8g, mentha camphor 0.15g, microencapsulation apple flavour 0.4g, matrix 10.0g, Walocel MT 20.000PV 5.0g, by total amount 1 kilogram, then add base-material.
Embodiment four
The preparation of IgG antibody toothpaste.Get IgG antibody 0.1g of the present invention, potassium sorbate 0.015g, Sodium Benzoate 0.015g, glycerine 10.0g, Sorbitol Powder 8.0g, Walocel MT 20.000PV 2.0g, Thallus Gracilariae 1.3g, Sodium Lauryl Sulphate BP/USP 1.8g mentha camphor 0.015g, aspartame 0.015g, strawberry flavour 0.05ml, calcium phosphate two water thing 47.8g.Add in the distilled water of about 60ml by Walocel MT 20.000PV, Thallus Gracilariae, swelling is dissolved, more successively adds other composition, fully stirs, makes to mix, finally add distilled water and be settled to 100ml, be stirred into paste.
Embodiment five
IgG antibody protects the preparation of tooth cream.Get IgG antibody 0.1g of the present invention, Sodium Benzoate 0.01g, beeswax 8.0g, stearic acid 10.0g Zerol 2.0g, glycerine 10.0g, Walocel MT 20.000PV 1.0g, mentha camphor 0.01g, aspartame 0.05g, distilled water 68.80ml, strawberry flavour 0.02g.By beeswax, stearic acid, Zerol, glycerine mixing, be heated to 70 DEG C, the liquid of mixed dissolution is referred to as A liquid; Walocel MT 20.000PV is dissolved in the distilled water of 50ml, swelling is dissolved, successively add IgG antibody, mentha camphor, aspartame, potassium sorbate, strawberry flavour again, abundant stirring, mix, then add cooled A liquid, finally add distilled water and be settled to 100ml, be stirred into paste.
Claims (8)
1. a preparation method for anti-periodontopathy bacteria antibody, is characterized in that comprising the steps:
(1) porphyromonas gingivalis is got, Anaerobic culturel 5-7 days in BHI substratum; Cultured porphyromonas gingivalis is carried out centrifugal treating, collects thalline, wash thalline 4-6 time with the phosphate buffer normal saline that pH6-7, concentration are 0.05-0.2M, at the temperature of 50-65 DEG C, heating 25-35 minute, carries out inactivation treatment, obtains porphyromonas gingivalis liquid;
The freund adjuvant of porphyromonas gingivalis liquid and equivalent is mixed, uses homogenizer process, obtained thalline adjuvant antigen;
(2), according to dried meat breast class animal species, get porphyromonas gingivalis antigen 1-5ml at every turn, lymphoglandula and dorsal sc multi-point injection are carried out to animal, inject three times altogether, every minor tick three weeks; After final immunization 20 days, to tire after testing qualified rear collection serum, get under immune serum stirs and add the dilution of appropriate salt solution, adjust ph, to 7.5-8.5, adds equivalent saturated ammonium sulphate solution, at 2-8 DEG C of temperature, leave standstill 20-30 hour, high speed centrifugation 20-30 minute, getting its precipitation, through melting ultrafiltration and concentration desalination, degerming postlyophilization again, obtaining resisting porphyromonas gingivalis IgG antibody crude extract;
(3), by after resisting porphyromonas gingivalis IgG antibody crude extract centrifugal purification, then through DEAE-SephadexA50 column chromatography; Carry out continuously or fraction gradient wash-out with the phosphoric acid buffer containing NaCl, collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, Function protein content is 20mg/ml;
By the protein eluate of previous step gained again through SephadexG200 gel filtration chromatography purifying, carry out wash-out with the phosphoric acid buffer containing NaCl, collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, degerming through 0.22 μ membrane filtration, lyophilize, obtains resisting porphyromonas gingivalis IgG antibody sterling.
2. the preparation method of a kind of anti-periodontopathy bacteria antibody according to claim 1, it is characterized in that the speed of centrifugal purification is 5000-12500rpm, centrifugal number of times 2-5 time, in DEAE anion exchange chromatography, the concentration of elutriant phosphoric acid buffer is 0.003-0.02M, the concentration of contained NaCl is 0.03-0.1M, and gel filtration chromatography elutriant phosphate buffer density is 0.01-0.2M, and the concentration of contained NaCl is 0.1-0.5M; Affinity chromatography eluate concentration is 0.1-0.5M.
3. the preparation method of a kind of anti-periodontopathy bacteria antibody according to claim 1 and 2, is characterized in that:
Use BHI substratum, under 37 DEG C of temperature anaerobic conditions, quiescent culture porphyromonas gingivalis 5-7 days, by the porphyromonas gingivalis 4000rpm after cultivation, centrifugal 10 minutes, collects thalline; With pH value be 6.0, concentration is that the phosphate buffer normal saline of 0.2M washes thalline 5 times, heat inactivation process in 50 DEG C, 25 minutes, obtained porphyromonas gingivalis liquid; Get 2,000,000,000/ml bacterium liquid, mix with the freund adjuvant of equivalent, use homogenizer process, obtained porphyromonas gingivalis adjuvant antigen;
According to mammal species, get thalline adjuvant antigen 1-5ml at every turn, lymphoglandula and subcutaneous multi-point injection are carried out to animal, inject three times altogether, every minor tick three weeks;
Third time immune animal after 20 days, detection tire qualified after, collect antiserum(antisera) salt solution to dilute, adjust ph to 8.0, add under stirring equivalent saturated ammonium sulphate solution put 2-8 DEG C leave standstill 24 hours, centrifugal 20 minutes of 8000rpm, gets precipitation, through ultrafiltration desalination, concentrated, degerming postlyophilization, obtained anti-periodontopathy bacteria antibody IgG crude extract;
By anti-periodontopathy bacteria antibody IgG crude extract through DEAE-SephadexA50 chromatography column chromatography; Post bed 2.5 × 35cm, adds the sample 3.0ml of 10mg/ml, carries out wash-out, flow velocity 20ml/h with 0.01M, pH7.0 phosphoric acid buffer containing 0.07MNaCl, and often pipe 5.0ml pressed by elutriant; Collect each albumen elution peak, and detect the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, Function protein content is 20mg/ml; Then by this liquid further through SephadexG200 gel filtration chromatography; Post bed 2.0 × 65cm, application of sample 1.5ml, carry out wash-out, flow velocity 8.0ml/h with 0.01M, pH7.0 phosphoric acid buffer containing 0.1MNaCl, and often pipe 5.0ml pressed by elutriant, collects each albumen elution peak, and detects the antibody activity of each albumen with ELISA; Collect the protein eluate having antibody activity, degerming through 0.22 μ membrane filtration, lyophilize, obtains resisting porphyromonas gingivalis IgG antibody sterling.
4. the application of the resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, it is characterized in that: to add in potassium sorbate, Sodium Benzoate two kinds of antiseptic-germicides at least one as the anti-periodontopathy composition of effective ingredient, comprise toothpaste, suck liquid, the oral product of collutory and aerosol, or chewing gum, chocolate, ice cream, ice cream food, the add-on of IgG antibody of the present invention is 0.05-0.2%, and the add-on of potassium sorbate, Sodium Benzoate is 0.01-0.02%.
5. the application of the resisting porphyromonas gingivalis IgG antibody adopting the inventive method to prepare, is characterized in that: the method for liquid is sucked in preparation, is get IgG antibody 2.0g of the present invention, potassium sorbate 0.15g, Sodium Benzoate 0.15g, aspartame 0.8g, mentha camphor 0.15g, apple flavour 0.4ml; Mentha camphor added in the distilled water of 100ml, 60 DEG C dissolve; Other solid content is respectively dissolved in the distilled water of 450ml, is then mixed by the uniform liquid of various dissolving, with distilled water constant volume to 1000ml, to obtain final product.
6. the application of the resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, it is characterized in that: the method preparing chewing gum, get IgG antibody 2.0g of the present invention, potassium sorbate 0.15g, aspartame 0.8g, mentha camphor 0.15g, microencapsulation apple flavour 0.4g, matrix 10.0g, Walocel MT 20.000PV 5.0g, by total amount 1 kilogram, to obtain final product.
7. the application of the resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, it is characterized in that: the method for toothpaste processed, get IgG antibody 0.1g of the present invention, potassium sorbate 0.015g, Sodium Benzoate 0.015g, glycerine 10.0g, Sorbitol Powder 8.0g, Walocel MT 20.000PV 2.0g, Thallus Gracilariae 1.3g, Sodium Lauryl Sulphate BP/USP 1.8g mentha camphor 0.015g, aspartame 0.015g, strawberry flavour 0.05ml, calcium phosphate two water thing 47.8g, by Walocel MT 20.000PV, Thallus Gracilariae adds in the distilled water of about 60ml, swelling is dissolved, successively add other composition again, abundant stirring, make to mix, finally add distilled water and be settled to 100ml, be stirred into paste, obtain.
8. the application of the resisting porphyromonas gingivalis IgG antibody prepared by the inventive method, it is characterized in that: the method for tooth cream is protected in preparation, get IgG antibody 0.1g of the present invention, Sodium Benzoate 0.01g, beeswax 8.0g, stearic acid 10.0g Zerol 2.0g, glycerine 10.0g, Walocel MT 20.000PV 1.0g, mentha camphor 0.01g, aspartame 0.05g, distilled water 68.80ml, strawberry flavour 0.02g, by beeswax, stearic acid, Zerol, glycerine mixing, be heated to 70 DEG C, the liquid of mixed dissolution is referred to as A liquid; Walocel MT 20.000PV is dissolved in the distilled water of 50ml, swelling is dissolved, successively add IgG antibody, mentha camphor, aspartame, potassium sorbate, strawberry flavour again, abundant stirring, mix, then add cooled A liquid, finally add distilled water and be settled to 100ml, be stirred into paste, obtain final product.
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