CN103520719B - A kind of buccal antibody product and preparation method thereof - Google Patents
A kind of buccal antibody product and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of buccal antibody product and preparation method thereof, described buccal antibody product contains safe and effective edible antibody.At least one virus following and/or antibacterial are had specificity to neutralize or inhibitory action by described antibody: influenza virus includes bird flu virus, respiratory syncytial virus, human parainfluenza viruses 3, streptococcus pneumoniae, hemophilus influenza, rhinovirus, coronavirus, rotavirus, adenovirus, enterovirus, respiratory syncytial virus or A group B streptococcus.Buccal antibody product of the present invention has the advantages such as low cost, high specificity, activity height, safety edible use, can be used for oral cavity or the cleaning of respiratory tract or the prevention of infectious disease or auxiliary treatment.
Description
Technical field
The present invention relates to biological technical field, particularly to a kind of buccal antibody product and preparation method thereof.
Background technology
Antibody drug is that target antigen (such as virus, antibacterial, cancerous cell etc.) is had the most special identification
With a kind of protein medicaments of rejection ability, owing to desired specificities is become by force by it to health side effect is little
The most potential newtype drug, is also that (price is to treat mesh equally to the most expensive medicine at present
Tens times of chemical drugs).Although antibody drug price is high but market golden eggs enriches, therefore, closely
The R&D direction that over Nian, each big pharmacy giant in the antibody drug Cheng Liao world chases.
The goods of sucking having prevention function of diseases the most on the market have Traditional-Chinese-medicine-type, fruit-vegetable type, vitamin type still
There are no buccal antibody product.Report one is had to can be used for the infectiousness epidemic disease disease epidemic prevention such as prevention of atypical pneumonia
Buccal tablet (number of patent application: CN03108028.6), it be a kind of with Radix Glycyrrhizae, Radix Angelicae Dahuricae (Radix Heraclei Scabridi), Herba Asari,
The raw material such as Borneolum Syntheticum, Cinnabaris be basic prescription be processed into suck goods.Another reported a kind of antiviral mouth
Buccal tablet (number of patent application: CN200610055156.5), it is a kind of with Chinese patent medicine antivirus oral liquid
It is that after major ingredient adds adjuvant, tabletted sucks goods that prescription and preparation technology extract acquisition crude extract.
Recently, Heilongjiang Institute of Agricultural Sciences's herding institute discloses a kind of extraction avian influenza egg yolk antibodies
Method (number of patent application: CN201110447572.0).Another reported (patent application
Number: CN02103760.4) disclose a kind of vaccine kept off infection by the inoculation mankind to the side of hen
Method, makes hen produce the egg containing specific immunity antibody.This immune egg the most edible is expected to eliminate
Internal inflammation and raising pest resistance.But, how above-mentioned report is not directed to, including with what shape
Formula, becomes the real effectively prophylactic proper product of energy by antibody producing.
Summary of the invention
The technical problem to be solved is to provide can be to being unfavorable for that health has containing one or more
Oral cavity or Respirovirus and antibacterial are especially had specificity to neutralize or the antibody of inhibitory action by pest matter,
Can be used for oral cavity or the cleaning of respiratory tract or the prevention of infectious disease or auxiliary treatment.
In order to solve above-mentioned technical problem, the technical solution used in the present invention is: a kind of buccal antibody product,
Including excipient, it is characterized in that having specificity neutralize or press down virus and/or antibacterial possibly together with at least one
The antibody made.
One or more viruses following and/or antibacterial are had specificity to neutralize or inhibitory action by described antibody.Institute
State virus and/or antibacterial include but not limited to: influenza virus includes bird flu virus, respiratory syncytial virus,
Human parainfluenza viruses 3, streptococcus pneumoniae, hemophilus influenza, rhinovirus, coronavirus, rotavirus,
Adenovirus, enterovirus, respiratory syncytial virus or A group B streptococcus.
In described buccal antibody product, final antibody titer is 100-100000 titre, preferably at 1000-10000
Titre scope.
Described antibody is to cross suckling mother animal (such as: milch cow and milk goat) from immunity, under Ruzhong extract
And at least one virus and/or antibacterial are had specificity neutralize or the active antibodies of inhibitory action.
Described antibody is the female bird (such as: hen) from immunity, under egg in extract and at least one
Plant virus and/or antibacterial has specificity and neutralizes or the active antibodies of inhibitory action.
Described antibody be the blood crossing animal from immunity extract and at least one virus and/or antibacterial
There is specificity neutralize or the active antibodies of inhibitory action.
Described antibody is to extract from cell culture fluid animal ascites and viral and/or thin at least one
Bacterium has specificity and neutralizes or the reactive monoclonal antibodies of inhibitory action.
Described antibody is to be obtained after immune animal by corresponding antigen.Described antigen can be one single
Viral or the combination lysate of multiple Strain or the strain alive of attenuation；Can also be the vaccine being commercially used,
Such as influenza A H1N1 vaccine；The selection of influenza virus can use the World Health Organization (WHO) to recommend
Seasonal current Influenza Virus；Can be the protein of any virus or antibacterial, especially virus or antibacterial be divided
The surface separated out or memebrane protein；Antigen protein can be prepared by technique for gene engineering or synthetic
The virus relevant with described respiratory tract infection or bacterioprotein overall or prepare according to its aminoacid sequence
Fragment or polypeptide；Described antigen can also be a kind of vaccine based on DNA, and it can be in immune animal body
Produce antigen.
The preparation method of a kind of buccal antibody product, including following several steps: 1) selection of antigen and preparation；
2) animal immune；3) antibody extracts；4) suck goods to prepare.
Antigen in described step 1) be select one or more viruses following and/or lysates of antibacterial or
The strain alive of attenuation, or the virus of purification or antibacterial associated protein, with Freund's complete adjuvant before immune animal
Or incomplete Freund's adjuvant mixes, emulsifying prepares；Described virus or antibacterial include but not limited to: influenza is sick
Poison, bird flu virus, respiratory syncytial virus, human parainfluenza viruses 3, streptococcus pneumoniae, and influenza is bloodthirsty
Bacillus, rhinovirus, coronavirus, rotavirus, adenovirus, enterovirus, respiratory syncytial virus or A
Group B streptococcus.
Described step 4) suck in goods preparation process, antibody or blended the raw material of antibody, half become
Product, finished product use less than 65 DEG C cryogenic systems.Described antibody need to be adjusted in the titer finally sucked in goods
100-100000 titre, preferably in 1000-10000 titre scope.
The technological process of described step 4) includes: 1) will suck product base crush, be dried, sieve standby
With；2) take the base material after part is sieved fully to mix, then adjust with purified water and the antibody-solutions got ready
With individually granulation, cold drying, broken, sieving for standby；3) by the base material after remaining sieving and other
Adjuvant and correctives fully mix the blank grain of system, dry, broken, sieving for standby；4) two kinds of granules are filled
Divide mixing, add tabletting after fluidizer, mix homogeneously, prepare antibody buccal tablet.
The technological process of described step 4) also includes: 1) gum base and syrup are dissolved at high temperature, are mixed；2)
Adding Icing Sugar and spice adjuvant, mixing makes micelle；3) micelle temperature is downgraded at 45-50 DEG C, add
Antibody activity composition, the most mixed；4) roll, molding of cutting into slices, make antibody chewing gum.
The invention has the beneficial effects as follows: antibody contained by buccal antibody product of the present invention is safe having
Imitate is edible.By disease prevention type buccal antibody product of the present invention, there is low cost, special
Property the advantage such as strong, activity height, safety edible use, can be used for oral cavity or the cleaning of respiratory tract or infectivity disease
Sick prevention or auxiliary treatment.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention is described in further detail:
The buccal antibody product of the present invention, including excipient, possibly together with at least one to virus and/or antibacterial
Specificity is had to neutralize or the antibody of inhibitory action.
One or more viruses following and/or antibacterial are had specificity to neutralize or inhibitory action by described antibody: stream
Influenza Virus, respiratory syncytial virus, human parainfluenza viruses 3, streptococcus pneumoniae, hemophilus influenza, nose
Virus, coronavirus, rotavirus, adenovirus, enterovirus, respiratory syncytial virus or A group B streptococcus.
In described buccal antibody product, final antibody titer is 100-100000 titre, preferably at 1000-10000
Titre scope.
Described antibody be extract from Ruzhong that immunity is crossed suckling mother animal and at least one virus and/
Or antibacterial has specificity and neutralizes or the active antibodies of inhibitory action.
Described antibody is the female bird from immunity, under egg in extract and at least one virus and/or
Antibacterial has specificity and neutralizes or the active antibodies of inhibitory action.
Described antibody be the blood crossing animal from immunity extract and at least one virus and/or antibacterial
There is specificity neutralize or the active antibodies of inhibitory action.
Described antibody is to extract from cell culture fluid animal ascites and viral and/or thin at least one
Bacterium has specificity and neutralizes or the reactive monoclonal antibodies of inhibitory action.
The preparation method of buccal antibody product, including following several steps: 1) selection of antigen and preparation；2)
Animal immune；3) antibody extracts；4) suck goods to prepare.
Antigen in described step 1) is to select one or more following virus and/or cracking of antibacterial
Liquid or the strain alive of attenuation, or the virus of purification or antibacterial associated protein, complete with Freund before immune animal
Full adjuvant (initial immunity is used) or incomplete Freund's adjuvant (follow-up immunization is used) mix, emulsifying is prepared；
Described virus or antibacterial include: influenza virus, bird flu virus (influenza virus), breathe
Road syncytial virus (RSV) (respiratory syncytial virus), human parainfluenza virus 3,
HPIV-3(human parainfluenza virus3), streptococcus pneumoniae (Streptococcus
Pneumoniae), hemophilus influenza (Haemophilus influenzae), such as: influenza B addicted to
Blood bacillus (Hib), rhinovirus, coronavirus (rhinovirus), rotavirus (coronavirus),
Adenovirus (adenovirus), enterovirus (enterovirus), respiratory syncytial virus
(respiratory syncytial virus), A group B streptococcus (group A streptococcus)
Deng.
Described step 4) suck in goods preparation process, antibody or blended the raw material of antibody, half become
Product, finished product use less than 65 DEG C, preferably less than 45 DEG C cryogenic systems.The technique stream of described step 4)
Journey includes: 1) will suck product base, and such as: soluble starch, sucrose etc., crush, is dried, sieves
Standby；2) take the base material after part is sieved fully to mix, then with purified water and the antibody-solutions got ready
Mediation individually granulation, cold drying, broken, sieving for standby；3) base material after remaining point being sieved with
Other adjuvant, such as: mannitol, and correctives, such as: Herba Menthae, citric acid, the blank grain of fully mixing system,
Dry, broken, sieving for standby；4) two kinds of granules are fully mixed, add fluidizer, such as: stearic acid
Tabletting after magnesium, mix homogeneously, makes antibody buccal tablet.
Described sucking goods and can also make gum formats, its preparation method is essentially: first by gum base
Dissolve at high temperature with syrup, mix, be subsequently adding the adjuvant such as Icing Sugar and spice, after mixing, micelle temperature is adjusted
Be down at 45-50 DEG C, now add antibody isoreactivity composition roll after fully mixing, molding of cutting into slices.Mouthful
The solvent ratio of fragrant sugar is: gum base 18-20%, sugar about 60%, glucose syrup about 20%, glycerol
0.3-0.5%, antibody is appropriate, and aromatizing agent and flavouring agent are appropriate.
Prepared by embodiment 1. influenza antigen:
H1N1, H3N2 influenza virus and B-mode Brisbane virus are cracked respectively then by 1:1:1
Ratio mixes, and three kinds of influenza virus cracking mixed liquors of system are standby.Immunity time, then by this mixed liquor with
Freund's complete adjuvant (for the first time immunity with) or press 1:1 with incomplete Freund's adjuvant (follow-up immunization with)
Ratio mixing, emulsifying prepare the antigen mixed liquor containing total antigen about 0.6mg/ml.
Prepared by embodiment 2. milk antibody:
The conceived milch cow (holstein cow) of choosing, at farrowing nine weeks, subcutaneous injection 50 milligrams a few days ago
Embodiment 1 described in the antigen mixed liquor of Freund's complete adjuvant emulsifying, carry out first immunisation, three
Week, with after seven weeks, mixes with the antigen using incomplete Freund's adjuvant emulsifying described in 30 milligrams of embodiments 1
It is each the most once that liquid distinguishes immunity again.After childbirth, then immunity double injection lymph node near thymus again.
Colostrum and often breast five months are collected continuously after animal childbirth.With 12,000g is centrifuged 15 minutes and removes butterfat
Fat, obtains defat antibody breast, through 60 DEG C of 30 minutes pasteurizations (notes: pasteurization can make antibody activity
Declining about 20%) subpackage, freezen protective are standby afterwards.Take a small amount of sample, exempt from for every kind in detection milk
The concentration of epidemic disease Globulins: use rabbit anti-cattle IgG, IgA and the IgM specific the of HRP coupling respectively
Two antibody and ELISA algoscopy carry out quantitative IgG, IgA and IgM concentration.This laboratory test results: immunity
The mean concentration of IgG, IgA and IgM in milk respectively reaches 0.3g/L, 0.02g/L and 0.01
g/L；Merge between rabbit anti-cattle IgG, IgA and the IgM specific second antibody warp of use HRP coupling
Connect ELISA method (this detection method is known method, can refer to the detection of antibody activity described in embodiment 4)
Detection, this antibody breast to H1N1, H3N2 and B-mode Brisbane three kinds of antigens of virus all in specific positive,
Titre reaches 600 times respectively, 450 times and 400 times.This antibody breast lyophilization is become lyophilized powder, subpackage,
Freezen protective is standby.
Prepared by embodiment 3 yolk antibody:
Influenza A H1N1 influenza virus and H influenzae type B (Hib) are cracked respectively then by 1:1
Ratio mixes, and prepares the antigen mixed liquor containing total antigen about 0.4mg/ml, standby.Choose primiparity or high yield
The healthy Rome hen of phase, to every hen by subcutaneous multi-point injection containing 0.2 milligram of antigen by 1:1
Ratio mixes with Freund's complete adjuvant, the antigen mixed liquor of emulsifying, carries out first immunisation.Pass through after two weeks
Injection is mixed containing the antigen of the mixing with incomplete Freund's adjuvant in 1:1 ratio of 0.1 milligram of antigen, emulsifying again
Close liquid, carry out secondary immunity.After two weeks by inject again injection again containing 0.1 milligram of antigen by 1:1
Ratio mixes with incomplete Freund's adjuvant, the antigen mixed liquor of emulsifying, carries out three immunity.It is collected from
Primary immune response plays the egg after 6 weeks.Inject the most again once containing 0.1 milligram of antigen by 1:1
Ratio mixes with incomplete Freund's adjuvant, the antigen mixed liquor of emulsifying.
From egg, isolate egg yolk, then add the deionized water of 5 times of volumes and adjust pH to 5.5, quiet
Put overnight, centrifuging and taking supernatant, then add PEG-6000 to 12%, room temperature is placed 30 minutes, 2000rpm
Centrifugal 15 minutes, gained precipitation was with the Tris-HCl(pH8.0 of the 0.01mol/L of 3 times of volumes) buffering
Liquid dissolves, it is thus achieved that anti-H1N1 influenza virus and the special yolk antibody liquid of anti-Hib hemophilus influenza.So
After, (this detection method is known method, can join to take yolk antibody liquid 0.1 milliliter indirect elisa method
Detect according to antibody activity described in embodiment 4) detection antibody titer, according to testing result, low temperature is dense
Contracting or with the Tris-HCl(pH8.0 of 0.01mol/L) buffer dilution tune gained antibody liquid is to titre extremely
More than 10000, then aseptic filtration, subpackage, freezen protective are standby.
Prepared by embodiment 4. antibody buccal tablet
1. antibody buccal tablet production technology:
A: by 1000 grams of soluble starch vacuum dehydrating at lower temperature, sieving for standby；100 grams of sucrose are pulverized,
80 mesh sieving for standby；
B: take soluble starch 200 grams and 20 grams of abundant mixings of cane sugar powder that part is sieved, mixing respectively
Afterwards with 10 grams of adjuvant mannitol, the titer got ready equal to or more than the antibody molten contracting liquid of 100000 titres
15ml(or antibody lyophilized powder add appropriate amount of purified water and reconcile into 10ml) pelletize, less than 45 DEG C cryogenic vacuums
Being dried, broken, 30 mesh sieve for subsequent use；
C: the remaining soluble starch sugarcane sieved and Icing Sugar are added 50 grams of mannitol and citric acid 1 gram
Fully mixing, the blank grain of system, it is dried, broken, 30 mesh sieve for subsequent use；
D: fully mixed by two kinds of granules, adds fluidizer magnesium stearate 10 grams, always mixes, mix homogeneously
Rear tabletting, every 1 gram；
E: quality testing；
F: aluminum-plastic packaged.
Note: for protect antibody activity, above-mentioned suck goods preparation technology must avoid high temperature (less than 65 DEG C,
Best less than 45 DEG C).
2. antibody activity detection:
Take a piece of buccal antibody product (1 gram) prepared by milk antibody and be soaked in the PBS of 5ml
Buffer, room temperature 1 hour, then within 10 minutes, it is centrifuged through 6000RPM rotating speed, obtains supernatant.
Antibody specificity measures: use indirect enzyme-linked immunosorbent detection method to measure, by H1N1 virus antigen mark
Quasi-product are (from United Kingdom National biological standard and the antigen cultivated by egg of control institute
California/7/2009 (NYMC X-179A), (NIBSC code: 09/146)) use PBS
Being diluted to 4ug/ml, add to by the amount of every groove 100ul in the reactive tank of enzyme bidding quotation, it is little that room temperature stands 3
Discard this liquid time after, and with PBS dishwashing 3 times, in each groove, then add 200ml envelope
Close after liquid (being dissolved in 1% human albumin HAS of PBS) room temperature stands 3 hours and discard this liquid,
Again with PBS dishwashing 3 times, airing is standby.Testing sample supernatant PBS is pressed 1:3
Gradient dilution, prepares 8 parts of different diluents, and a copy of it does not contains any testing sample, every part of 0.2ml;
Every part takes 100ul and adds in grooves different on the enzyme bidding quotation of above-mentioned antigen coated mistake, and every part of sample surveys two
Secondary, room temperature stationary incubation discards this liquid after 2 hours, with cleaning mixture, (0.1%Tween20 is dissolved in PBS
Buffer) dishwashing 3 times；Take that the anti-cattle of rabbit two of 100ul HRP labelling is anti-(to be carried when sucking antibody in goods
When taking from milk) or the dilution of rabbit anti-chicken IgY bis-anti-(when sucking that in goods, antibody extracts from egg)
Liquid (0.05% is dissolved in the HBS buffer containing 1% human albumin) adds in each groove of enzyme bidding quotation,
Room temperature stationary incubation discards this liquid after 1 hour, with cleaning mixture, (0.1%Tween20 is dissolved in PBS buffering
Liquid) dishwashing 3 times；Take in each groove that the 100ul TMB nitrite ion containing H2O2 adds enzyme bidding quotation to, room
Gentle and quiet putting occurs to blue gradient, then adds the H2SO4 stop buffer of 100ul1M to each of enzyme bidding quotation
Color development stopping in groove；The absorbance of each groove is detected at 450 nm by microplate reader.
Testing result (such as following table) display is surveyed and is sucked goods specific antibody activity relatively by force, and specificity resists
Body titre surpasses 1200.
Table 1, goods Specific antibody titre result is sucked in Dot-ELISA detection
Prepared by embodiment 5. antibody chewing gum
The Radix Acaciae senegalis of 4kg is dissolved at 80 DEG C softening 2 hours, 4kg glucose is dissolved in 2kg
Water in boil to 107 DEG C, then with Radix Acaciae senegalis stirring mixing, add the fine sugar that sieves of 12kg
Powder (granularity is between 0.1mm mono-0.05mm) and 60 milliliters of glycerol, stirring mixing, downgrade micelle
Temperature is between 45 50 DEG C；Take antibody molten contracting liquid or antibody lyophilized powder adds pure water and is adjusted to 200 milliliters
Titer is 10000-100000 titre (preparation method for antibody is shown in above-described embodiment), joins in micelle,
Till being fully blended into uniformly；The micelle mixed is cut into every piece of about 4kg, and at 30-60 minute
Inside it is allowed to cool, then rolls, cut into slices, Icing Sugar Yu layer, packaging.
The Efficacy experiments that flu-prevention is infected by embodiment 6. buccal antibody product
Selecting 20 families of volunteer, the condition of selecting is: there are 3 people, Fu Muhe in each volunteer family
One 10 years old or following child's (regardless of sex), wherein child has suffered from influenza and has obvious influenza disease
Shape, father and mother do not have any flu-like symptom, father and mother to live together with suffering from influenza child.
The volunteer family of select is randomly divided into two groups: matched group and antibody group, often group 10
Family.To each family of matched group father and mother and suffer from influenza child's buccal without antibody suck goods, give
The father and mother of each family of antibody group and suffer from influenza child's buccal influenza antibodies and suck goods, daytime is little every 3
Time a piece of to buccal mouth, every day 5, use 30 days continuously, observe and record father and mother's sense in each family
The situation of dye influenza.
Result of the test is as shown in the table: have 7 people to infect influenza, infection rate 35% in matched group father and mother；
Antibody group father and mother there are 4 people infect influenza, infection rate 20%.Obviously, buccal antibody product has necessarily
Flu-prevention effect.
Embodiment described above is merely to illustrate technological thought and the feature of the present invention, its object is to make
Those skilled in the art are it will be appreciated that present disclosure implementing according to this, it is impossible to only with the present embodiment
Limiting the scope of the claims of the present invention, what the most all disclosed spirit was made changes on an equal basis or modifies,
Still fall in the scope of the claims of the present invention.
Claims (2)
1. the preparation method of a buccal antibody product, it is characterized in that, including following several steps: 1) selection of antigen and preparation: described antigen is the combination of influenza A H1N1 influenza virus and H influenzae type B, influenza A H1N1 influenza virus and H influenzae type B are cracked respectively and then mixes in 1:1 ratio, prepare the hybrid antigen liquid containing total antigen 0.4mg/ml；2) animal immune and antibody extract: for the animal immune of antigen mixed liquor and the concretely comprising the following steps of antibody extraction of influenza A H1N1 influenza virus and H influenzae type B: choosing the healthy Rome hen of primiparity or high yield period, the antigen mixed liquor in 1:1 ratio Yu Freund's complete adjuvant mixing and emulsifying that by subcutaneous multi-point injection, every hen is contained 0.2 milligram of antigen carries out first immunisation；After two weeks, the injection antigen mixed liquor in 1:1 ratio Yu incomplete Freund's adjuvant mixing and emulsifying containing 0.1 milligram of antigen carries out secondary immunity again, by the injection antigen mixed liquor in 1:1 ratio Yu incomplete Freund's adjuvant mixing and emulsifying containing 0.1 milligram of antigen again after two weeks, carry out three immunity；It is collected from the egg after for the first time immunity rises 6 weeks, injects the most again once containing the mixing with incomplete Freund's adjuvant in 1:1 ratio of 0.1 milligram of antigen, the antigen mixed liquor of emulsifying；From egg, isolate egg yolk, then add the deionized water of 5 times of volumes and adjust pH to 5.5, standing overnight；Centrifuging and taking supernatant, then PEG-6000 to 12% is added, room temperature is placed 30 minutes, 2000rpm is centrifuged 15 minutes, retain precipitation, gained precipitation is with the Tris-HCl buffer solution of 0.01mol/L, pH8.0 of 3 times of volumes, just yolk antibody liquid is obtained, described yolk antibody liquid 0.1ml indirect elisa method is detected antibody titer, according to testing result cryoconcentration or dilute with the Tris-HCl buffer of 0.01mol/L, pH8.0 that to adjust gained antibody liquid to titre be more than 10000, then aseptic filtration, subpackage, freezen protective are standby；3) suck goods to prepare: the soluble starch of 1000g carries out vacuum dehydrating at lower temperature, sieving for standby, is pulverized by 100g sucrose, 80 mesh sieving for standby；Take the soluble starch 200g partly sieved and cane sugar powder 20g respectively fully to mix, add, equal to or more than the antibody molten contracting liquid 15ml of 100000 titres or antibody lyophilized powder, the 10ml mixed liquor that appropriate amount of purified water reconciles into pelletize with adjuvant mannitol 10g and the titer got ready after mixing, crush again after less than 45 DEG C low-temperature vacuum dryings, 30 mesh sieve for subsequent use；Remaining sieve soluble starch and cane sugar powder are fully mixed plus the citric acid of mannitol 50g and 1g, make blank grain, be dried, broken, 30 mesh sieve for subsequent use；Two kinds of granules are fully mixed, adds fluidizer magnesium stearate 10g, always mix, tabletting after mix homogeneously, make antibody buccal tablet, every 1g.
2. the preparation method of a buccal antibody product, it is characterized in that, including following several steps: 1) selection of antigen and preparation: described antigen is H1N1, H3N2 influenza virus and the combination of B-mode Brisbane virus, then its preparation method is mixed in 1:1:1 ratio for H1N1, H3N2 influenza virus and B-mode Brisbane virus being cracked respectively, prepare three kinds of influenza virus cracking mixed liquors, standby；Mixed in 1:1 ratio with Freund's complete adjuvant or with incomplete Freund's adjuvant by this mixed liquor during immunity, emulsifying prepares the antigen mixed liquor containing total antigen 0.6mg/ml again；2) animal immune and antibody extract: for concretely comprising the following steps that H1N1, H3N2 influenza virus and the animal immune of B-mode Brisbane virus antigen mixed liquor and antibody extract: select a conceived holstein cow, in farrowing nine week a few days ago, at the antigen mixed liquor of the Freund's complete adjuvant emulsifying of its subcutaneous injection 50 milligrams, carry out first immunisation；After three weeks and seven weeks, then with 30 milligrams of antigen mixed liquors through incomplete Freund's adjuvant emulsifying, distinguish immunity each the most once；After childbirth, then the lymph node injection near thymus twice；Colostrum and often breast five months are collected continuously after animal childbirth；It is centrifuged 15 minutes with 12000g and removes butter oil；Gained defat antibody breast subpackage, freezen protective after 60 DEG C of 30 minutes pasteurizations is standby, take the concentration of every kind of immunoglobulins in a small amount of sample detection milk, respectively with the rabbit anti-cattle IgG of HRP coupling, IgA, the specific second antibody of IgM and ELISA algoscopy carry out quantitative IgG, IgA, IgM concentration；3) suck goods to prepare: 4000g Radix Acaciae senegalis is dissolved at 80 DEG C softening 2 hours, 4000g glucose is dissolved in the water of 2000g and boils to 107 DEG C, then with Radix Acaciae senegalis stirring mixing, add the ultrafine powdered sugar that granularity is 0.1-0.05mm and the glycerol of 60ml that 12kg sieves, stirring mixing, downgrades micelle temperature between 45-50 DEG C；Taking antibody molten contracting liquid or antibody lyophilized powder adds pure water and is adjusted to 200ml, titer is 10000-100000 titre, joins in micelle, till being sufficiently mixed to uniformly；The micelle mixed is cut into every piece for 4kg, and be allowed to cool in 30-60 minute, then roll, cut into slices, Icing Sugar Yu layer, packaging, make antibody chewing gum.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1557339A (en) * | 2004-01-15 | 2004-12-29 | 高春平 | Preparation for preventing and treating tympanitis and dental caries |
| CN1613344A (en) * | 2004-09-17 | 2005-05-11 | 珠海百奥生物技术有限公司 | Anti-dental caries chewing gum and preparation |
| CN1680445A (en) * | 2005-01-12 | 2005-10-12 | 江西3L医用制品集团有限公司 | Anti-hemolytic streptococcus, staphylococcus and candida albicans 1gG antibody and preparation method |
| CN101564534A (en) * | 2008-11-12 | 2009-10-28 | 大连理工大学 | Specific yolk antibody preparation for controlling halitosis pathogens and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0920684A (en) * | 1995-07-07 | 1997-01-21 | Lotte Co Ltd | Chewing gum |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1557339A (en) * | 2004-01-15 | 2004-12-29 | 高春平 | Preparation for preventing and treating tympanitis and dental caries |
| CN1613344A (en) * | 2004-09-17 | 2005-05-11 | 珠海百奥生物技术有限公司 | Anti-dental caries chewing gum and preparation |
| CN1680445A (en) * | 2005-01-12 | 2005-10-12 | 江西3L医用制品集团有限公司 | Anti-hemolytic streptococcus, staphylococcus and candida albicans 1gG antibody and preparation method |
| CN101564534A (en) * | 2008-11-12 | 2009-10-28 | 大连理工大学 | Specific yolk antibody preparation for controlling halitosis pathogens and application thereof |
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