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Method for inactivating viruses during avian influenza H9 subtype vaccine production

A technology for the production of avian influenza viruses and vaccines, applied in biochemical equipment and methods, vaccines, viruses, etc., can solve the problems of destroying the immunogenicity of pathogens and long time for formaldehyde inactivation

Pending Publication Date: 2020-09-29
ZHAOQING DAHUANONG BIOLOGIC PHARMA
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

In the production process of domestic bird flu inactivated vaccines, the inactivation process uses formaldehyde solution as an inactivator, and its inactivation can achieve good results, but formaldehyde inactivation not only inactivates the virus but also destroys the immunity of pathogens The inactivation time of formaldehyde is relatively long, and it is easily affected by factors such as temperature, pH value, concentration, presence of organic matter, type of pathogen, and nitrogen content. Residual free formaldehyde, if injected into the body with the vaccine, will cause adverse reactions irritative reaction

Method used

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  • Method for inactivating viruses during avian influenza H9 subtype vaccine production
  • Method for inactivating viruses during avian influenza H9 subtype vaccine production
  • Method for inactivating viruses during avian influenza H9 subtype vaccine production

Examples

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Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: Optimization of inactivation process conditions

[0019] Test material: H9N2 subtype avian influenza virus

[0020] Inactivation method: (1) Virus inoculation and cultivation: select LMH cells for vaccine preparation and subculture until the density of LMH cells reaches 1.0×10 6 -1.5×10 6 Each / mL was inoculated with avian influenza virus H9 subtype, and added to cell maintenance medium for propagation and culture; (2) Virus collection, concentration and purification: the cell culture medium in step (1) was frozen and thawed three times, and stored at 4°C, After centrifugation at 7000rpm for 15 minutes, collect the avian influenza virus H9 subtype supernatant, and ultrafilter and concentrate the above supernatants respectively to obtain the avian influenza virus H9 subtype virus venom; (3) Virus inactivation: in the negative pressure production area, According to the amount of virus liquid, add BPL with final concentrations of 0.005%, 0.015%, 0.025%, 0.05...

Embodiment 2

[0032] Embodiment 2: vaccine preparation

[0033] Vaccine preparation: Mix 94 parts of white oil, 6 parts of Tween, and 1.5 parts of aluminum stearate to prepare the oil phase, mix and emulsify the solution of avian influenza virus and the oil phase according to the volume ratio of 1:2, and prepare the H9 subtype of avian influenza vaccine.

[0034] Table 3: Vaccines prepared by different inactivation processes

[0035]

Embodiment 4

[0036] Embodiment 4: BPL inactivated vaccine safety test

[0037] Test method: 60 SPF chickens were randomly divided into 6 groups, 10 birds in each group, among which 5 batches of BPL inactivated vaccine were used for safety evaluation in groups 1-5, and 2.5 mL was injected intramuscularly into the chest of each chicken. 6 groups of blank control group, no inoculation.

[0038] Safety test results: 14 days after immunization, the chickens in each treatment group were healthy, and the vaccine was well absorbed by necropsy, and no abnormalities appeared.

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Abstract

The invention provides a method for inactivating viruses during avian influenza H9 subtype vaccine production. By optimizing a virus inactivating process, the safety of vaccine preparations can be significantly improved, the stress response is reduced, and the immune effect is improved.

Description

technical field [0001] The invention belongs to the field of animal vaccines, in particular to a method for inactivating viruses in the production of avian influenza H9 subtype vaccines. Background technique [0002] According to different pathogenicity, avian influenza can be divided into low pathogenic avian influenza and highly pathogenic avian influenza. The highly sick avian influenza is represented by the H7N9 subtype, and the low pathogenic avian influenza is represented by the H9N2 subtype. H9N2 subtype avian influenza is a bird flu that spreads quickly and spreads widely in poultry in my country; chickens of various ages are susceptible, and the clinical severity varies greatly, but they all cause great economic losses. Egg production drops by 10%-60% or is completely eliminated, and the death rate of broiler chickens and brooding or rearing chickens ranges from 10% to 80%; Co-infection with various immunosuppressive pathogens, especially secondary infection with Es...

Claims

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Application Information

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IPC IPC(8): C12N7/06A61K39/145A61P31/16
CPCC12N7/00A61K39/12A61P31/16C12N2760/16163C12N2760/16134A61K2039/5252A61K2039/552
Inventor 张文炎刘之文詹烜子刘锦容高艳张云丽王土活
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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