CN109045295A - A kind of application of vaccine adjuvant and preparation method thereof - Google Patents
A kind of application of vaccine adjuvant and preparation method thereof Download PDFInfo
- Publication number
- CN109045295A CN109045295A CN201811049533.3A CN201811049533A CN109045295A CN 109045295 A CN109045295 A CN 109045295A CN 201811049533 A CN201811049533 A CN 201811049533A CN 109045295 A CN109045295 A CN 109045295A
- Authority
- CN
- China
- Prior art keywords
- biochanin
- water
- vaccine adjuvant
- soluble
- vaccine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses applications of a kind of vaccine adjuvant and preparation method thereof, using Biochanin A or water-soluble Biochanin A as a kind of vaccine adjuvant.Preparation method is the following steps are included: Biochanin A or water-soluble Biochanin A are dissolved in physiological saline, 0.22 μm of membrane filtration degerming；Water-soluble Biochanin A it is water-soluble the preparation method comprises the following steps: Biochanin A is completely dissolved with 70% ethanol solution after, NaOH is added and adjusts pH to 9-13, the heating water bath 4-6h in 50-70 degrees Celsius of water-bath, dry to constant weight, the water-soluble Biochanin A after being modified is concentrated under reduced pressure.Application of the invention is that obtained Biochanin A or water-soluble Biochanin A are had the function of beasts immunostimulant as beasts vaccine adjuvant, which is immunoreacted, easy to use, safe, without toxicity.
Description
Technical field
The present invention relates to a kind of veterinary biologics technical fields, and in particular to a kind of application and its preparation of vaccine adjuvant
Method.
Background technique
Vaccine immunity is one of the important means of prevention and control zoonosis, and vaccine has used many subunit antigens in recent years
And polypeptide antigen, such antigen has the characteristics that purity is high, safety are good, but immunogenicity is poor, cannot effectively induce and exempt from
Epidemic disease reaction, therefore improve its immunogenicity using adjuvant and be of great significance.Have not applied to the vaccine adjuvant of livestock and poultry at present
Family name's adjuvant, aluminium hydroxide etc., although the immune effect of vaccine can be improved in such adjuvant, in the safety used there is also
It is insufficient.Compared with synthetic drug, Chinese medicine has the characteristics that pleiotropism, two-way immune regulative, adverse reaction is few, by tcm development
Being developed into safely and effectively novel vaccine adjuvant is a kind of new approach, is had broad application prospects.
Biochanin A (BiochaninA, BioA) belongs to isoflavone compound, is Chinese medical extract, is primarily present
In the leguminous herbaceous plants such as red clover, chick-pea, have the function of antiviral and antitumor and anti-inflammatory.Research finds olecranon
Bean sprout vegetarian A inhibits the expression of NLRP3 to mitigate the hepatic injury of LPS induction by activation Nrf2 approach；BioA passes through increased SOD, GPx
Concentration reduces MDA content to protect cerebral neuron；Biochanin A inhibits LPS to induce peripheral blood mononuclear cells NF- κ B
Activation, adjusts the secretion of cell factor.Biochanin A is not easy to absorb in vivo, is led to bioavilability due to poorly water-soluble
It is low, its drug effect and utilization are affected, there is not been reported for the vaccine adjuvant effect in relation to Biochanin A at present.
There is patent CN102018143A in the prior art, function is immunized using the feed regulating yellow broiler containing isoflavones
The method of energy and muscle antioxygenic property；It is disclosed that a kind of broiler fodder containing isoflavones is provided using the feed
Chicken body production performance, immunity and chicken meat quality；Its working substance is isoflavones, and effect body is broiler chicken, and by adding
Forage feed is added to take, the maximum effect of isoflavones can not be fully achieved by the absorption of forage feed, and
Other type functions of osajin also need to be researched and developed.
Summary of the invention
In view of the problems of the existing technology, the present invention provides a kind of application and preparation method thereof of vaccine adjuvant, this is answered
It is immunoreacted with enhancing, it is easy to use, safe, without toxicity.
Application of a kind of vaccine adjuvant and preparation method thereof, including following content:
A kind of application of vaccine adjuvant, which is characterized in that using Biochanin A or water-soluble Biochanin A as one
Kind vaccine adjuvant.
Further, the Biochanin A or water-soluble Biochanin A are as a kind of live vaccine adjuvant.
Further, the assistant of the Biochanin A or water-soluble Biochanin A as oralbumin vaccine
Agent.
Further, the vaccine adjuvant is that the physiology salt that Biochanin A concentration is 50-200 μ g/100 μ l is water-soluble
Liquid, or the normal saline solution for being 12.5-100 μ g/100 μ l for water-soluble Biochanin A concentration.
Further, the vaccine adjuvant and vaccine are injected after 1:1 is mixed by volume and are used.
Further, the vaccine adjuvant storage temperature is 2-6 DEG C.
A kind of preparation method of vaccine adjuvant, which is characterized in that Biochanin A or water-soluble Biochanin A is molten
Solution is in physiological saline, with vaccine adjuvant obtained after membrane filtration degerming.
Further, the water-soluble Biochanin A is water-soluble the preparation method comprises the following steps: pressing mass parts for 1 part of olecranon
After bean sprout vegetarian A is completely dissolved with 70% ethanol solution of 8-20 part, NaOH is added and adjusts pH to 9-13, in 50-70 degrees Celsius of water
Heating water bath 4-6h in bath is dried under reduced pressure to constant weight, the water-soluble Biochanin A after being modified.
The beneficial effects of the present invention are as follows:
1, for live vaccine adjuvant provided by the invention to animal without toxicity, good immune effect efficiently solves vaccine
The problem of side reaction；Wherein, Biochanin A Biochanin A more unmodified after Na ion chelating increases water solubility
Identical vaccine adjuvant effect is played when lower concentration, and mouse spleen lymphocyte breeder reaction can be improved, improve in serum specifically
Property IgG antibody it is horizontal.
2, dosage form good water solubility easy to use, that live vaccine adjuvant of the invention obtains, solution are reduced without lamination
Existing aluminium glue adjuvant will carry out pre-temperature, when the temperature is excessively high using the preceding use technology for needing to shake up when environment temperature is lower
It is easily layered, lamination is then not present in the present invention；
3, vdiverse in function, adjuvant of the present invention not only has the function of similar aluminium glue adjuvant sustained release antigen, also has significant increase
The function of strong humoral immunity of organism and cellular immunity；
4, ingredient is simple, and preparation process is simple, and equipment requirement is low, easily stores；Development and utilization in livestock and poultry breeding industry are latent
Power is huge, has broad application prospects.
Detailed description of the invention
Fig. 1 is immunoglobulin G (IgG) test result of various concentration adjuvant；
Fig. 2 is the test result of splenic lymphocyte proliferation rate under different antigenic actions；
Specific embodiment
The present invention is further described with reference to embodiments.
Embodiment 1
Application of a kind of vaccine adjuvant and preparation method thereof, steps are as follows:
(1) 0.5-2g Biochanin A is dissolved in 1000ml physiological saline, 0.22 μm of membrane filtration degerming obtains
Concentration is the Biochanin A normal saline solution of 50-200 μ g/100 μ l, and preservation temperature is 4 DEG C.(2) by 0.1ml step
(2) Biochanin A normal saline solution is mixed with 0.1ml oralbumin vaccine, and being made has immune thorn
The vaccine for swashing function, is injected into beasts body and is immunized.
Embodiment 2
Application of a kind of vaccine adjuvant and preparation method thereof, steps are as follows:
(1) the water-soluble modification of Biochanin A
After 2g Biochanin A is dissolved with 20ml70% ethyl alcohol, it is 9-13 that NaoH adjustment pH, which is added, Celsius in 50-70
Heating water bath 4-6h in the water-bath of degree is dried under reduced pressure to constant weight to get the water-soluble Biochanin A to after modifying, water
Dissolubility improves 263-382 times；
(2) preparation of vaccine adjuvant
The water-soluble Biochanin A in 0.125-2g step (1) is taken to be dissolved in 1000ml physiological saline, 0.22 μm of filter
Film filtration sterilization, is prepared into 12.5-200 μ g/100 μ L water solubility Biochanin A vaccine adjuvant, and storage temperature is 4 DEG C.
(3) by the water-soluble Biochanin A normal saline solution and 0.1ml oralbumin in 0.1ml step (2)
Vaccine mixing, is made the vaccine with immunostimulation function, is injected into beasts body and is immunized.
Test result display cooperation 0.1ml vaccine uses 0.1ml Biochanin A or water-soluble Biochanin A physiology
When saline solution is as adjuvant, adjuvant of the Biochanin A content in 50 μ g/100 μ l has started effectiveness, water-soluble chickpea sprout
Adjuvant of the plain A content in 12.5 μ g/100 μ l has started effectiveness, and as concentration increases, effectiveness will increase, and theoretically may be selected
The Biochanin A of nearly 100% concentration, effectiveness is best, and selecting the concentration to 200 μ g/100 μ l is the peace for conventional dose
Full property is considered.
Water-soluble Biochanin A (abbreviation wBioA used below) and Biochanin A (abbreviation BioA used below) are exempted from
The experiment of epidemic disease adjuvanticity
Material and method: being randomly divided into 10 groups (6/group) for Female ICR mice, and it is as follows that 10 experimental group difference are arranged:
(1) oralbumin (abbreviation OVA used below) control group: every abdominal part hypodermic OVA solution (1mg/mL)
0.2mL；
(2) aluminium hydroxide control group: every abdominal part hypodermic 0.2mL contains the OVA solution (1mg/ of 200 μ g aluminium hydroxides
mL)；
(3) wBioA experimental group: every injection 0.2mL contains the OVA solution (1mg/ of wBioA (12.5,25,50,100 μ g)
mL)；
(4) BioA experimental group: every injection 0.2mL contains the OVA solution (1mg/mL) of BioA (12.5,25,50,100 μ g)；
Each group immune 2 times, first immunisation and secondary immunity interval two weeks.
Two exempt from rear two weeks execution mouse, and acquisition promotees blood coagulation, separates serum, and -80 DEG C of refrigerators save backup；Sterile removal spleen
It is dirty, prepare spleen lymphocyte suspension.
It is immune using specific antibody immunoglobulin G (IgG) potency in ELISA method, CCK-8 method detection serum and OVA
Mouse spleen lymphocyte breeder reaction.
Referring to attached drawing 1, peripheral blood serum IgG Concentration Testing interpretation of result is as follows:
Two exempt from after 50 and 100 μ g BioA group-specific IgG antibodies it is horizontal significant or it is extremely significant be higher than OVA group (P < 0.05 or
P<0.01)；12.5 μ g wBioA group IgG antibody levels are significantly higher than OVA group (P < 0.05), 25 μ g wBioA and higher concentration group
IgG antibody is horizontal extremely significant higher than OVA group (P < 0.01), and 25 μ g wBioA groups and 100 μ g BioA group IgG antibody levels exist
Same level；There are significant difference (P < 0.05) between the IgG antibody concentration of the BioA and wBioA group of every kind of concentration.
Referring to attached drawing 2, splenic lymphocyte proliferation rate situation has:
WBioA or BioA as after adjuvant and OVA co-immunization mouse, spleen lymphocyte in vitro by ConA, LPS and
The lymphopoiesis ability that OVA stimulation generates is significant or extremely significant higher than OVA group (P < 0.05 or P < 0.01)；WBioA adjuvant
Lymphocyte proliferation rate has raising trend after being increased to 50 μ g by 25 μ g of concentration, but the indifference opposite sex changes；With BioA adjuvant group phase
Than wBioA adjuvant group is remarkably improved ConA and LPS induction splenic lymphocyte proliferation rate (P < 0.05).
The result shows that: wBioA can significantly improve OVA specific IgG antibody titer in immune serum, improve ConA,
Mouse lymphocyte breeder reaction is immunized in the OVA of LPS and OVA induction；Compared with BioA, 25 μ g wBioA adjuvant groups induce OVA
Specific IgG antibody titer and 100 μ g BioA adjuvant group levels are thought together in immune serum, 25, the induction of 50wBioA μ g group
Stimulation lymphopoiesis ability is significantly higher than BioA group.
Claims (8)
1. a kind of application of vaccine adjuvant, which is characterized in that using Biochanin A or water-soluble Biochanin A as one kind
Vaccine adjuvant.
2. a kind of application of vaccine adjuvant according to claim 1, which is characterized in that by the Biochanin A or
Water-soluble Biochanin A is as a kind of live vaccine adjuvant.
3. a kind of application of vaccine adjuvant according to claim 1, which is characterized in that by the Biochanin A or
Adjuvant of the water-soluble Biochanin A as oralbumin vaccine.
4. the application of any vaccine adjuvant according to claim 1 to 3, which is characterized in that the vaccine adjuvant is hawk
Mouth bean sprout vegetarian A concentration is the normal saline solution of 50-200 μ g/100 μ l, or is that water-soluble Biochanin A concentration is
12.5-200 the normal saline solution of μ g/100 μ l.
5. a kind of application of vaccine adjuvant according to claim 4, which is characterized in that the vaccine adjuvant is pressed with vaccine
It injects and uses after volume ratio 1:1 mixing.
6. the application of any vaccine adjuvant described in -5 according to claim 1, which is characterized in that the vaccine adjuvant saves
Temperature is 2-6 DEG C.
7. a kind of preparation method of vaccine adjuvant, which is characterized in that by Biochanin A or water-soluble Biochanin A dissolution
In physiological saline, with vaccine adjuvant obtained after membrane filtration degerming.
8. a kind of preparation method of vaccine adjuvant according to claim 7, which is characterized in that the water-soluble chick-pea
Bud element A is water-soluble the preparation method comprises the following steps: being completely dissolved 1 part of Biochanin A with 70% ethanol solution of 8-20 part by mass parts
Afterwards, NaOH is added and adjusts pH to 9-13, the heating water bath 4-6h in 50-70 degrees Celsius of water-bath, be concentrated under reduced pressure dry to perseverance
Weight, the water-soluble Biochanin A after being modified.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811049533.3A CN109045295A (en) | 2018-09-10 | 2018-09-10 | A kind of application of vaccine adjuvant and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811049533.3A CN109045295A (en) | 2018-09-10 | 2018-09-10 | A kind of application of vaccine adjuvant and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109045295A true CN109045295A (en) | 2018-12-21 |
Family
ID=64761075
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811049533.3A Pending CN109045295A (en) | 2018-09-10 | 2018-09-10 | A kind of application of vaccine adjuvant and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109045295A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110339353A (en) * | 2019-06-18 | 2019-10-18 | 龙岩学院 | Application of the red clover as haemophilus parasuis vaccine adjuvant |
| CN114306590A (en) * | 2021-12-31 | 2022-04-12 | 皖西学院 | Veterinary vaccine diluent with immunity enhancement effect and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080226682A1 (en) * | 2007-03-13 | 2008-09-18 | Peros Systems Technologies, Inc. | Immunoactive Compositions for Improved Oral Delivery of Vaccines and Therapeutic Agents |
| CN102373248A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Method for purifying biochanin A |
-
2018
- 2018-09-10 CN CN201811049533.3A patent/CN109045295A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080226682A1 (en) * | 2007-03-13 | 2008-09-18 | Peros Systems Technologies, Inc. | Immunoactive Compositions for Improved Oral Delivery of Vaccines and Therapeutic Agents |
| CN102373248A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Method for purifying biochanin A |
Non-Patent Citations (2)
| Title |
|---|
| 郝鹏飞等: "《天然药物化学》", 31 July 2014, 吉林大学出版社 * |
| 陈洪博等: "红车轴草异黄酮免疫佐剂作用的研究", 《 中国畜牧兽医学会兽医病理学分会八届五次常务理事会议暨第二十二次学术研讨会、中国病理生理学会动物病理生理学专业委员会学术研讨会》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110339353A (en) * | 2019-06-18 | 2019-10-18 | 龙岩学院 | Application of the red clover as haemophilus parasuis vaccine adjuvant |
| CN114306590A (en) * | 2021-12-31 | 2022-04-12 | 皖西学院 | Veterinary vaccine diluent with immunity enhancement effect and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105198988B (en) | Anti- Vibrio splindidus Yolk antibody and preparation method thereof | |
| CN105324114B (en) | Astaxanthin anti-inflammatory synergistic combination | |
| Qiu et al. | Immunopotentiating effects of four Chinese herbal polysaccharides administered at vaccination in chickens | |
| Chen et al. | The immune-adjuvant activity and the mechanism of resveratrol on pseudorabies virus vaccine in a mouse model | |
| EP1557171A1 (en) | Rabbit skin comprising biological active substance and its use | |
| CN109045295A (en) | A kind of application of vaccine adjuvant and preparation method thereof | |
| Yang et al. | The Taishan Robinia pseudoacacia polysaccharides enhance immune effects of rabbit haemorrhagic disease virus inactivated vaccines | |
| CN107158087A (en) | It is a kind of to prevent Chinese medicine composition of hog cholera and preparation method thereof | |
| CN105343032B (en) | A kind of Fenbendazole micro-capsule and preparation method thereof | |
| CN101259160B (en) | Compound preparations for treating birdsvirosis and preparation thereof | |
| CN103564495A (en) | Propolis and cordycepic hypha powder composition, as well as preparation and application thereof | |
| CN101816663B (en) | Medicinal composition and injection for curing avian toxoplasmosis, and preparation method thereof | |
| CN108926710A (en) | A kind of animal enteric oral gel vaccine and its preparation, application method | |
| CN103393680A (en) | Application of berberine in medicine for treating non-alcoholic fatty liver disease | |
| CN103520316B (en) | Traditional Chinese medicine for preventing and treating livestock and poultry diarrhoea and preparation method | |
| CN103446182B (en) | Preparation method of specific transfer factor of highly pathogenic porcine reproductive and respiratory syndrome | |
| CN101647788B (en) | Florfenicol microcapsule and preparation method thereof | |
| CN107056959A (en) | Jerusalem artichoke moderate resistance HSV 1, the composition of RSV, EV 71 and preparation | |
| CN104208683B (en) | The pharmaceutical composition of a kind of anti-duck viral disease, lyophilized powder, preparation method and application | |
| CN109078055B (en) | Isoferulic acid, traditional Chinese medicine extract containing isoferulic acid and application of cimicifuga foetida | |
| CN106176634A (en) | A kind of transmissible gastroenteritis of swine epidemic diarrhea Bi-combined attenuated Vaccine liposome dilution freeze-dried products and preparation method thereof | |
| CN105106950B (en) | A kind of immunologic adjuvant of live vaccine, preparation method and applications | |
| Miao et al. | Immuno-Enhancing Effect of Ginsenoside Rh2 Liposomes on Foot-and-Mouth Disease Vaccine | |
| CN104784152B (en) | A kind of Thiamphenicol micro-capsule and preparation method and application | |
| CN105031245B (en) | A kind of Chinese medicine composition of anti-Eimeria Tenella |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181221 |
|
| RJ01 | Rejection of invention patent application after publication |