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A kind of preparation method of h7n9 recombinant influenza virus inactivated vaccine

A technology for influenza virus and inactivated vaccines, which is applied in the field of preparation of H7N9 recombinant influenza virus inactivated vaccines, can solve the problems of multi-epitope mucosal immunization vaccines without good curative effect, and achieve great application value, strong immunogenicity and low cost low effect

Active Publication Date: 2019-04-19
ZHAOQING DAHUANONG BIOLOGIC PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In terms of vaccine research against avian influenza H7N9 virus, there is no mucosal immune vaccine with good efficacy against H7N9 subtype in the prior art, and there is no multi-epitope mucosal immune vaccine with good efficacy against H7N9 subtype virus. The epidemic of possible avian influenza H7N9 virus infection, there is an urgent need for a new and highly effective vaccine to prevent avian influenza

Method used

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  • A kind of preparation method of h7n9 recombinant influenza virus inactivated vaccine
  • A kind of preparation method of h7n9 recombinant influenza virus inactivated vaccine
  • A kind of preparation method of h7n9 recombinant influenza virus inactivated vaccine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation method of the avian influenza virus split vaccine of the present embodiment comprises the steps of: inoculating 10-day-old SPF chicken embryos with H7N9 avian influenza recombinant virus, establishing the main generation seed batch virus seeds of avian influenza, and starting from the main generation seed batch of avian influenza Among the virus seeds, the virus seeds with high hemagglutination titer were selected as the working seeds to batch establish the virus seeds.

[0022] The above-mentioned obtained H7N9 avian influenza working seed batch virus species was diluted to 10 with sterilized PBS. -5 Dilution, inoculated into the allantoic cavity of 10-day-old healthy SPF chicken embryos, the virus inoculation volume was 0.1ml / embryo, and the chicken embryos were cultured at 35°C for 40 hours; live embryos were selected and placed in cold embryos at 4°C for 15 hours, and the virus allantois was harvested liquid.

[0023] Add 0.1% formaldehyde to the ob...

Embodiment 2

[0029] The preparation method of avian influenza virus split vaccine comprises the following steps: inoculating 10-day-old SPF chicken embryos with H7N9 avian influenza recombinant virus, establishing a batch of avian influenza main generation seed batch virus seeds, and selecting from the avian influenza main generation seed batch virus seeds Virus seeds with high hemagglutination titer are used as working seeds to batch establish virus seeds.

[0030] The above-mentioned obtained H7N9 avian influenza working seed batch virus species was diluted to 10 with sterilized PBS. -7 Dilution, inoculated into the allantoic cavity of 10-day-old healthy SPF chicken embryos, the virus inoculation volume was 0.1ml / embryo, and the chicken embryos were cultured at 37°C for 72 hours; live embryos were selected and placed in cold embryos at 4°C for 20 hours, and the virus allantois was harvested liquid.

[0031] Add 0.1% formaldehyde to the obtained viral allantoic fluid and inactivate it at...

Embodiment 3

[0036] Embodiment three: experiment report.

[0037] In this experiment, the existing platform of the laboratory was used to rapidly prepare the H7N9 avian influenza vaccine, and immunized SPF chickens to observe the effect of the vaccine.

[0038] 2.1 Materials

[0039] 2.1.1 Molecular cloning related materials

[0040] 2.2.1.1 Strains: Top10 E. Coli, BJ5183 E. coli, XL-1Blue E. Coli competent cells.

[0041] 2.2.1.2 E1E3, pGA1 (provided by Dr. Peter Palease) Δ plasmid: pAd-

[0042] 2.2.1.3 HA7 and NA9 genes

[0043] The HA7 gene and NA9 gene of influenza virus A / Anhui / 1 / 2013 (H7N9) were obtained from the GISAID influenza database and synthesized with reference to the database sequence.

[0044] 2.2.1.4 Culture materials: sterile petri dish, LB, SOB medium and corresponding solid medium preparation methods refer to "Molecular Cloning Experiment Guide".

[0045] 2.1.2 Reagents: various restriction enzymes and T4 DNA ligase were purchased from Takara and NEB; various Taq ...

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Abstract

The invention discloses a preparation method of an inactivated H7N9 recombinant influenza virus vaccine. The steps include the establishment of batch virus seeds of avian influenza working seeds; virus inoculation and cultivation; virus inactivation; virus harvest liquid filtration and concentration; virus purification; sugar removal; It is prepared as a semi-finished product of H7N9 avian influenza virus inactivated vaccine. The H7N9 recombinant influenza inactivated vaccine prepared according to the method of the invention has strong immunogenicity, broad antigen spectrum, stable quality, high purity, good safety, low cost, is suitable for large-scale production, and has great application value.

Description

technical field [0001] The invention belongs to the field of vaccine preparation, and in particular relates to a preparation method of H7N9 recombinant influenza virus inactivated vaccine. Background technique [0002] Influenza is the abbreviation of influenza. It is a zoonotic infectious disease caused by influenza virus. The main symptoms after human infection are high fever, cough, runny nose, myalgia, etc. Most of them are accompanied by severe pneumonia. The failure of various organs such as kidneys and kidneys leads to death, and the case fatality rate is very high; the H7N9 subtype influenza that has occurred since April 2013 has posed a huge threat to humans and poultry. At present, vaccines are mainly used for prevention, and the vaccines used include virus inactivated vaccines, attenuated vaccines, and recombinant subunit vaccines. [0003] The outer membrane of influenza virus particles is covered by two types of surface glycoproteins, one type is hemagglutinin ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/145C12N7/00A61P31/16C12R1/93
Inventor 罗琴芳潘蔚绮冯立强陈凌陈瑞爱钟南山
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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