CN107287268A - A kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1 - Google Patents
A kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1 Download PDFInfo
- Publication number
- CN107287268A CN107287268A CN201710457600.4A CN201710457600A CN107287268A CN 107287268 A CN107287268 A CN 107287268A CN 201710457600 A CN201710457600 A CN 201710457600A CN 107287268 A CN107287268 A CN 107287268A
- Authority
- CN
- China
- Prior art keywords
- enzymolysis
- nucleic acid
- hiv
- polypeptide according
- enzymolysis polypeptide
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- 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
- A61K39/12—Viral antigens
-
- 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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
-
- 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
- A61K2039/55516—Proteins; Peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16211—Human Immunodeficiency Virus, HIV concerning HIV gagpol
- C12N2740/16234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Immunology (AREA)
- Public Health (AREA)
- General Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Virology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1, the adjuvant is a kind of enzymolysis polypeptide, is prepared via a method which to form：Using sheep bone as the substrate of enzyme digestion reaction, with trypsin digestion, with the 3000u of ultrafiltration retaining molecular weight 5000 component, freeze-drying.Enzymolysis polypeptide of the present invention can significantly increase the immunogenicity of HIV nucleic acid vaccines, strengthen the vivo immunization reply efficiency of HIV nucleic acid vaccines, be favorably improved the immune effect of HIV nucleic acid vaccines.
Description
Technical field
The invention belongs to field is immunized, and in particular to a kind of adjuvant of enhancing HIV-1 nucleic acid vaccine immunity originality.
Background technology
DNA vaccination has the advantages that simple, economic, the easily stored transport of making, makes it in bacterium, virus, parasite etc.
There is very big potential using value in the preventing and treating and treatment of a variety of diseases, may be to the preventing and treating of human diseases and being good for for animal husbandry
Kang Fazhan plays epoch-making effect.U.S. FAD approved hepatitis B vaccines etc. more than tens plant DNA vaccination and enter clinical test, show
The great potential and application prospect of DNA vaccination are shown.And due to the weaker reason of the immunogenicity of DNA vaccination, often will be with assistant
Agent is used in conjunction with strengthening its action effect.
AIDS (AIDS) is to be entered by AIDS virus (HIV) after body, destroys the immunocyte of body, ultimately results in
The immunocompetence of body is wholly absent, so as to trigger a kind of serious infectious disease of various diseases.Encode HIV retrovirus
The gene of albumen mainly has three kinds：Env、Pol、Gag.The nuclear protein and reverse transcriptase egg of Pol and Gag main codes virus
In vain, and the albumen of Env gene main codes its viral membrane surfaces, and neutralizing antibody epitope substantially all on Env genes.Mesh
Before there is no effective vaccine and medicine to be used for the inhibition of HIV for preventing and removing infection.DNA vaccination has as emerging vaccine
Many advantages, such as traditional vaccine can not be replaced, both at home and abroad also regards DNA vaccination as in one of key technology for capturing AIDS, but mesh
The greatest problem of preceding restriction DNA vaccination development is that immunogenicity is not strong, therefore finds effective adjuvant to strengthen the effect of DNA vaccination
It is really extremely urgent.
The content of the invention
It is contemplated that overcome the defect of prior art strengthens HIV DNA epidemic diseases as adjuvant there is provided a kind of enzymolysis polypeptide
The immunogenicity of seedling, strengthens the vivo immunization reply efficiency of HIV DNA vaccinations.
The present invention is achieved by following technical solution：
A kind of enzymolysis polypeptide, is prepared via a method which to form：Using sheep bone as the substrate of enzyme digestion reaction, trypsase enzyme is used
Solution, with ultrafiltration retaining molecular weight 5000-3000u component, freeze-drying.
Preferably, enzymatic hydrolysis condition is pH value 7.8-8.2,46-50 DEG C of hydrolysis temperature, enzymolysis time 3-5 hours.
Preferably, the enzymolysis polypeptide is prepared via a method which to form：Fresh sheep bone is cleaned with clear water, through 0.08-
0.12MPa autoclavings 0.5-1.5h softens it, is broken into 1-3cm3Skeletal grain, be placed in distilled water, add trypsase
Enzymolysis；After enzymolysis terminates, heating inactivates trypsase, is cooled to normal temperature, 12000-14000 revs/min of 15-25 points of centrifugation
Clock, takes supernatant, carries out classification separation with the milipore filter that molecular cut off is 10000u, 5000u, 3000u successively, collects retention point
Son amount 5000-3000u component, freeze-drying.
Preferably, trypsase addition is the 0.3-0.5% of substrate weight.
Preferably, enzymatic hydrolysis condition is pH value 8.0,48 DEG C of hydrolysis temperature, enzymolysis time 4 hours.
Preferably, heating make trypsase inactivate method be：Water-bath 8-12 minutes under the conditions of 80-90 DEG C.
Preferably, soften it through 0.1MPa autoclavings 1h.
Above-mentioned enzymolysis polypeptide is used for the purposes of HIV-1 nucleic acid vaccine adjuvants.
Advantage of the present invention：
Enzymolysis polypeptide of the present invention can significantly increase the immunogenicity of HIV nucleic acid vaccines, strengthen the internal of HIV nucleic acid vaccines
Immune response efficiency (including humoral immunity and cellular immunity), is favorably improved the immune effect of HIV nucleic acid vaccines.
Brief description of the drawings
Fig. 1 is routine IgG elisa assay HIV-1Env specific humoral immunity response intensities.
Embodiment
In order to preferably explain technical scheme, it is further described with reference to specific embodiment.In embodiment
The experiment material do not emphasized especially is normal experiment material, belongs to the category that those skilled in the art are easily obtained.
Embodiment 1：The preparation of enzymolysis polypeptide
- 20 DEG C of freezen protectives are standby after fresh sheep bone clear water is cleaned；Ultrafiltration cup and milipore filter are purchased from Shanghai and rubbed fast science device
Material Co., Ltd；Trypsase (＞ 2.5 × 105U/g), Beijing Suo Laibao Science and Technology Ltd.
Enzymolysis polypeptide is prepared via a method which to form：
Fresh sheep bone is cleaned with clear water, softens it through 0.1MPa autoclavings 1h, is broken into 1-3cm3Skeletal grain, put
In distilled water, trypsin digestion (trypsase addition is the 0.4% of substrate weight) is added, enzymatic hydrolysis condition is pH value
8.0,48 DEG C of hydrolysis temperature, enzymolysis time 4 hours；After enzymolysis terminates, water-bath makes trypsase go out in 10 minutes under the conditions of 85 DEG C
It is living, be cooled to normal temperature, 13000 revs/min centrifuge 20 minutes, take supernatant, successively with molecular cut off be 10000u, 5000u,
3000u milipore filter carries out classification separation, collects molecular cut off 5000-3000u component, freeze-drying.
Embodiment 2：The immunological enhancement of enzymolysis polypeptide
First, experiment material
Enzymolysis polypeptide is prepared according to the method for embodiment 1.
SPF grades of female Balb/c mouse of 6-8 week old, for immunity evaluation.
2nd, experimental method
1st, the preparation of HIV-1 DNAs
HIV-1 plasmids carry out improved epidemic disease for the commission sharp rich structure in Guangzhou for HIV-1CN54 membranous antigens gp 145
Seedling plasmid, the genes of gp 145 are inserted in nucleic acid vaccine vector plasmid pVAX1, eukaryon expression plasmid pVAXGP is built into, has demonstrate,proved
In fact can high efficient expression.By pVAXGP vaccine plasmids and pVAX1 empty plasmids difference Transformed E .coli DH5a, plasmid is extracted, through enzyme
Sequencing identification is cut for positive colony, positive colony is inoculated in the LB fluid nutrient mediums containing ampicillin, shaken in 37 DEG C respectively
Culture 15h is swung, next day presses 1:100 expand the seed that culture 6h prepares fermented and cultured, are finally transferred to fermentation cylinder for fermentation culture, adopt
Concentration is adjusted to 1mg/mL in -20 DEG C of preservations with extensive alkaline lysis method of extracting plasmid, and with physiological saline.
2nd, animal packet and immunization method
Balb/c mouse are randomly divided into 3 groups, and every group 10, packet and processing method are as follows：
Empty plasmid group：Only immune pVAX1 empty plasmids, 50 μ L/ times/only；
Vaccine plasmid group：Only immune pVAXGP vaccine plasmids, 50 μ L/ times/only；
Adjuvant assisted group：Immune pVAXGP vaccine plasmids+enzymolysis polypeptide, 50 μ of pVAXGP vaccine plasmids L/ times/, enzymolysis
Polypeptide 2mg/kg/ times/.
When immune, muscle multi-point injection is in mouse both legs tibialis anterior.Respectively the 0th, 2,4 weeks immune mouse, at the 6th week
The dead every amynologic index of mouse detection.
3rd, amynologic index is detected
3.1INF- γ and IL-2ELISPOT detect specific cell immunoreaction
Splenic lymphocytes are obtained after putting to death mouse, with the CD8 of CN54Env albumen+T cell Dominant Epitopes peptide (amino acid sequence：
CKEVHNVWATHACVPTDPNP, SELYKYKVVEIKPLGIAPTA, QQSNLLRAIEAQQHLLQLTV) stimulate 24h, BD
The T lymphocytes formation spot number of Mouse IFN-γs and IL-2ELISPOT kits detection secretion of gamma-IFN and IL-2, to comment
Valency T lymphocyte specific immune response level.
3.2 binding antibody ELISA and affinity ELISA detection specific humoral immune responses
It is more than 95% HIV-1CN54 strain eukaryotic expression gp120 albumen coating elisa plate (0.01mg/L), 4 DEG C with purity
Overnight.Post-immunisation serum is with 1:100 be that initial dilution carries out doubling dilution, determines absorbance, titration is to terminal with calculating
Geometric mean titers.Affinity of antibody ELISA uses 0-5mol/L gradient NaSCN competition bindings after sera incubation under normal temperature
Antigen 1 5min, PBST washing 10 times, IgG-HRP secondary antibodies are incubated, washed afterwards, colour developing is equal to routine ELISA method.
3.3 statistical analysis
T inspections, P are carried out using SPSS20.0 statistics softwares<0.05 thinks significant difference.
3rd, experimental result
1st, specific cell immunoreaction
Put to death mouse and obtain splenic lymphocytes, utilize ELISPOT experimental analysis specific Cs D8+T cell immune response.
The experimental results are shown inthe following table by ELISPOT：
| Empty plasmid group | Vaccine plasmid group | Adjuvant assisted group | |
| SFC/106Cell (INF- γ) | ＜ 5 | 225 | 430 |
| SFC/106Cell (IL-2) | ＜ 5 | 36 | 85 |
Above table can be seen that enzymolysis polypeptide can significantly improve HIV-1 nucleic acid vaccines specific cellular immunity should
Answer.
2nd, specific humoral immune response
Immunized mice serum binding antibody ELISA and affinity ELISA detect specific humoral immune response, conventional
IgG elisa assay HIV-1Env specific humoral immunity response intensities are as shown in Figure 1；The experiment of NaSCN competitive ELISAs can divide
The change of antigen-specific IgG antibody affinity is analysed, 50% titre using IgG in initial serum is ED50Antibody parent can be evaluated
And power, each group affinity of antibody result is as shown in the table：
| Empty plasmid group | Vaccine plasmid group | Adjuvant assisted group | |
| ED50(mol/L) | 0.8 | 1.4 | 3.7 |
Can be seen that enzymolysis polypeptide from upper table and Fig. 1 can significantly improve the specific humoral immunity of HIV-1 nucleic acid vaccines
Response.
Enzymolysis polypeptide of the present invention can significantly increase the immunogenicity of HIV nucleic acid vaccines, strengthen the internal of HIV nucleic acid vaccines
Immune response efficiency, is favorably improved the immune effect of HIV nucleic acid vaccines.
Above-described embodiment is only used for that technical scheme is explained further, it will be appreciated by those skilled in the art that, appoint
What simple replacement is changed all without departing from the present invention, and protection scope of the present invention is not limited to above-mentioned specific embodiment.
Claims (8)
1. a kind of enzymolysis polypeptide, it is characterised in that be prepared via a method which to form：Using sheep bone as the substrate of enzyme digestion reaction, use
Trypsin digestion, with ultrafiltration retaining molecular weight 5000-3000u component, freeze-drying.
2. enzymolysis polypeptide according to claim 1, it is characterised in that：Enzymatic hydrolysis condition is pH value 7.8-8.2, hydrolysis temperature
46-50 DEG C, enzymolysis time 3-5 hours.
3. enzymolysis polypeptide according to claim 2, it is characterised in that be prepared via a method which to form：By fresh sheep bone
Cleaned with clear water, soften it through 0.08-0.12MPa autoclavings 0.5-1.5h, be broken into 1-3cm3Skeletal grain, be placed in distillation
In water, trypsin digestion is added；After enzymolysis terminates, heating inactivates trypsase, is cooled to normal temperature, and 12000-14000 turns/
Minute centrifugation 15-25 minutes, takes supernatant, is classified successively with the milipore filter that molecular cut off is 10000u, 5000u, 3000u
Separation, collects molecular cut off 5000-3000u component, freeze-drying.
4. enzymolysis polypeptide according to claim 3, it is characterised in that：Trypsase addition is substrate weight 0.3-
0.5%.
5. enzymolysis polypeptide according to claim 3, it is characterised in that：Enzymatic hydrolysis condition be pH value 8.0,48 DEG C of hydrolysis temperature,
Enzymolysis time 4 hours.
6. enzymolysis polypeptide according to claim 3, it is characterised in that the method that heating inactivates trypsase is：In 80-
Water-bath 8-12 minutes under the conditions of 90 DEG C.
7. enzymolysis polypeptide according to claim 3, it is characterised in that：Soften it through 0.1MPa autoclavings 1h.
8. any enzymolysis polypeptides of claim 1-7 are used for the purposes of HIV-1 nucleic acid vaccine adjuvants.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710457600.4A CN107287268A (en) | 2017-06-16 | 2017-06-16 | A kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710457600.4A CN107287268A (en) | 2017-06-16 | 2017-06-16 | A kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107287268A true CN107287268A (en) | 2017-10-24 |
Family
ID=60097046
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710457600.4A Pending CN107287268A (en) | 2017-06-16 | 2017-06-16 | A kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107287268A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109082453A (en) * | 2018-08-28 | 2018-12-25 | 青海七彩花生物科技有限公司 | A kind of polypeptide improves the purposes of influenza vaccines immune effect as influenza vaccines adjuvant |
| CN109097425A (en) * | 2018-08-28 | 2018-12-28 | 青海七彩花生物科技有限公司 | A kind of polypeptide and the purposes as influenza vaccines immunologic adjuvant |
| CN109182430A (en) * | 2018-08-28 | 2019-01-11 | 青海七彩花生物科技有限公司 | A kind of influenza vaccines immunologic adjuvant |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1327069A (en) * | 2001-05-24 | 2001-12-19 | 谢启安 | Process for extracting amino acid polypeptide with animal bone marrow and tissue |
| WO2002083216A1 (en) * | 2001-04-13 | 2002-10-24 | Becton Dickinson And Company | Method of intradermally injecting substances |
| CN101647815A (en) * | 2008-08-13 | 2010-02-17 | 北京凯瑞创新医药科技有限公司 | Bionic enzymatic product of animal medicament and application thereof |
| CN104651434A (en) * | 2015-01-28 | 2015-05-27 | 南京新百药业有限公司 | Preparation method of bone peptide solution |
| CN105876660A (en) * | 2016-04-25 | 2016-08-24 | 西南科技大学 | Method for preparing yak-sheep bone powder by utilizing enzyme method |
-
2017
- 2017-06-16 CN CN201710457600.4A patent/CN107287268A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002083216A1 (en) * | 2001-04-13 | 2002-10-24 | Becton Dickinson And Company | Method of intradermally injecting substances |
| CN1327069A (en) * | 2001-05-24 | 2001-12-19 | 谢启安 | Process for extracting amino acid polypeptide with animal bone marrow and tissue |
| CN101647815A (en) * | 2008-08-13 | 2010-02-17 | 北京凯瑞创新医药科技有限公司 | Bionic enzymatic product of animal medicament and application thereof |
| CN104651434A (en) * | 2015-01-28 | 2015-05-27 | 南京新百药业有限公司 | Preparation method of bone peptide solution |
| CN105876660A (en) * | 2016-04-25 | 2016-08-24 | 西南科技大学 | Method for preparing yak-sheep bone powder by utilizing enzyme method |
Non-Patent Citations (4)
| Title |
|---|
| 孙卫青等: "酶法水解鲜羊骨骼的研究", 《肉类研究》 * |
| 蔡丽华等: "骨蛋白酶解及风味肽分离现代技术研究进展", 《肉类研究》 * |
| 霍乃蕊等: "羊骨酶解物的免疫调节活性研究", 《核农学报》 * |
| 马俪珍等: "羊骨汤不同酶解方式对其成分的影响", 《食品研究与开发》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109082453A (en) * | 2018-08-28 | 2018-12-25 | 青海七彩花生物科技有限公司 | A kind of polypeptide improves the purposes of influenza vaccines immune effect as influenza vaccines adjuvant |
| CN109097425A (en) * | 2018-08-28 | 2018-12-28 | 青海七彩花生物科技有限公司 | A kind of polypeptide and the purposes as influenza vaccines immunologic adjuvant |
| CN109182430A (en) * | 2018-08-28 | 2019-01-11 | 青海七彩花生物科技有限公司 | A kind of influenza vaccines immunologic adjuvant |
| CN109097425B (en) * | 2018-08-28 | 2022-06-07 | 浙江康嘉基因技术有限公司 | Polypeptide and application of polypeptide as influenza vaccine immunologic adjuvant |
| CN109082453B (en) * | 2018-08-28 | 2022-10-21 | 南京赛尔健生物技术有限公司 | Application of polypeptide as influenza vaccine adjuvant in improving influenza vaccine immune effect |
| CN109182430B (en) * | 2018-08-28 | 2022-10-21 | 南京赛尔健生物技术有限公司 | Influenza vaccine immunologic adjuvant |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110760006B (en) | African swine fever immune system targeting genetic engineering vaccine | |
| CN107287268A (en) | A kind of adjuvant of the nucleic acid vaccine immunity originality of enhancing HIV 1 | |
| CN101885760A (en) | HIV-1gp120 gene consensus sequence optimized by codon and gp120 nucleic acid vaccine | |
| CN101062941A (en) | Recombined hepatitis E hepatitis virus protein, preparation method and usage thereof | |
| CN102221618B (en) | Method for establishing hog cholera lapinized virus labeled vaccine strain and preparing vaccine | |
| CN104873967A (en) | O type foot and mouth disease virus-like particle vaccine as well as preparation method and application thereof | |
| CN110981968B (en) | Fusion protein containing rabies virus G protein, preparation method, application and vaccine thereof | |
| CN104693310A (en) | Chimeric protein, virus-like particle and application thereof | |
| CN109136198A (en) | A kind of expression Chicken Infectious Anemia Virus VP1, VP2 genetic recombination bird pox virus live vector vaccine | |
| CN101457231A (en) | Construction of bacteria ghost vector PMAL-E-SN and its application in bacillus coli | |
| CN102210860B (en) | Mycobacterium tuberculosis TB10.4-F1 fusion protein vaccine and preparation method thereof | |
| CN102680699B (en) | ELISA (enzyme-linked immunosorbent assay) detection method for identifying fowl adenovirus group I (FAVI) infection | |
| CN104628865A (en) | Pseudorabies virus epitope polypeptide gene engineering vaccine | |
| CN103539839A (en) | Neutralizing epitope peptide of enterovirus 71-type VP2 antigen and application thereof | |
| CN109021115A (en) | A kind of pig circular ring virus trivalent subunit vaccine | |
| CN104974231A (en) | Novel porcine reproductive and respiratory syndrome virus variant GP5 recombinant protein and preparation method and application thereof | |
| CN102618557A (en) | Recombinant avian flavivirus E protein and application thereof | |
| CN103992408A (en) | Preparation of blue ear disease protein engineering vaccine | |
| CN102406929A (en) | Co-expressed molecular adjuvant enhanced divalent foot and mouth disease protein engineering vaccine | |
| CN115340609B (en) | Foot-and-mouth disease virus multi-antigen epitope fusion protein, protein cage nanoparticle and preparation method thereof | |
| CN101838319B (en) | Asia 1-type FMDV compound multiepitope | |
| CN103614329A (en) | Construction method and application of DNA (Deoxyribonucleic Acid) vaccine for avian leukosis virus subgroup J | |
| CN110484515B (en) | Vaccine vector for preventing FAdV-4 and NDV, and preparation method and application thereof | |
| CN113861277A (en) | Bovine rotavirus recombinant VP8 protein and application thereof | |
| CN107325160B (en) | Non-human primate cynomolgus monkey hepatitis B virus effector T cell specific antigen polypeptide and vaccine |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20171024 |
|
| WD01 | Invention patent application deemed withdrawn after publication |