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Preparation method of trivalent inactivated vaccine

A technology of inactivated vaccines and virus seeds, which is applied in the field of veterinary biological products, can solve the problems of low virus titer and high production cost, and achieve the effect of ensuring added and complete immune protection

Inactive Publication Date: 2017-11-03
广州博恒生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, avian adenovirus type 4 and avian reovirus type 2 are mainly propagated through SPF chicken embryo vectors, the virus titer is not high, and the production cost of large-scale production with SPF chicken embryo vectors is very expensive; Passage cell-derived cells to propagate the virus, and then use it to produce inactivated vaccines

Method used

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  • Preparation method of trivalent inactivated vaccine
  • Preparation method of trivalent inactivated vaccine
  • Preparation method of trivalent inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 A kind of preparation method of avian influenza virus, avian adenovirus and avian reovirus triple inactivated vaccine

[0032] The preparation method of described avian influenza virus, avian adenovirus and avian reovirus triple inactivated vaccine comprises the following steps:

[0033] S1. Preparation of cells: culture LMH cells for seedling production with serum-free cell growth medium until the density of LMH cells reaches 1.2×10 6 cells / mL, subcultured in a microcarrier bioreactor, wherein the amount of microcarriers used was 8.5g / L; the serum-free cell growth solution was composed of the following components and their contents: DMEM basic culture solution 85% v / v, ornithine 0.5mmol / L, behenic acid 0.55g / L, myristic acid 0.4g / L, guanosine 1.0mg / L, dihydrotanshinone I 5.5mg / L, lysine 4g / L, glycine 0.85g / L, phenylisoleucine 2.5g / L, arginine 3g / L, threonine 1.5g / L, valine 2.5g / L, transferrin 7.5g / mL, mugwort leaf polysaccharide 0.12g / L, pyridoxine hyd...

Embodiment 2

[0040] Embodiment 2 A kind of preparation method of avian influenza virus, avian adenovirus and avian reovirus triple inactivated vaccine

[0041] The preparation method of described avian influenza virus, avian adenovirus and avian reovirus triple inactivated vaccine comprises the following steps:

[0042] S1. Preparation of cells: use serum-free cell growth medium to cultivate LMH cells for seedling production until the density of LMH cells reaches 1.5×10 6 cells / mL for subculture; the serum-free cell growth solution is composed of the following components and their contents: 90% v / v of DMEM basic culture solution, 0.6mmol / L of ornithine, and 0.75g / L of behenic acid , myristic acid 0.5g / L, guanosine 1.5mg / L, dihydrotanshinone I 7mg / L, lysine 5.5g / L, glycine 1.0g / L, phenylisoleucine 3.5g / L, arginine Acid 5.0g / L, threonine 3g / L, valine 3.5g / L, transferrin 10g / mL, mugwort leaf polysaccharide 0.15g / L, pyridoxine hydrochloride 6mg / L, biotin 1.5mg / L, Zinc sulfate 0.8mg / L, sodium...

Embodiment 3

[0049] Embodiment 3 A kind of preparation method of avian influenza virus, avian adenovirus and avian reovirus triple inactivated vaccine

[0050] The preparation method of described avian influenza virus, avian adenovirus and avian reovirus triple inactivated vaccine comprises the following steps:

[0051] S1. Preparation of cells: use serum-free cell growth medium to culture LMH cells for seedling production until the density of LMH cells reaches 1.0×10 6cells / mL for subculture; the serum-free cell growth solution is composed of the following components and their contents: 90% v / v of DMEM base culture solution, 0.3mmol / L of ornithine, and 0.15g / L of behenic acid , myristic acid 0.3g / L, guanosine 0.5mg / L, dihydrotanshinone I 3.5mg / L, lysine 1.5g / L, glycine 0.75g / L, phenylisoleucine 1g / L, arginine Acid 2.0g / L, threonine 1g / L, valine 2g / L, transferrin 5g / mL, mugwort polysaccharide 0.05g / L, pyridoxine hydrochloride 3mg / L, biotin 0.1mg / L, sulfuric acid Zinc 0.3mg / L, sodium bica...

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Abstract

The invention belongs to the technical field of veterinary biological products and particularly relates to a preparation method of a trivalent inactivated vaccine for avian influenza virus, aviadenovirus and avian reovirus. The trivalent inactivated vaccine for avian influenza virus subtype H9, an aviadenovirus type 4 and avian reovirus type 2 is produced by adopting a virus high-adaptability continuous cell line LMH cell. The produced trivalent inactivated vaccine for the avian influenza virus subtype H9, the aviadenovirus type 4 and the avian reovirus type 2 is used for immunizing SPF (Specific Pathogen Free) chickens, so that a high-level HI (Hemagglutination Inhibiting) antibody for resisting the avian influenza virus subtype H9 and a high-level neutralizing antibody for resisting the aviadenovirus type 4 and the avian reovirus type 2 can be generated. The trivalent inactivated vaccine provided by the invention has high titer and complete immune protecting effect on the avian influenza virus subtype H9, the aviadenovirus type 4 and the avian reovirus type 2.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a preparation method of a triple inactivated vaccine of avian influenza virus, avian adenovirus and avian reovirus. Background technique [0002] H9N2 subtype avian influenza belongs to low pathogenicity avian influenza with high morbidity but low mortality. The main clinical manifestations are only mild respiratory symptoms, reduced feed intake, egg production rate can drop from 90% to below 20%, or even complete production, and about 30% of commercial broilers can die, and it is very easy to mix infection with Escherichia coli. Seriously affect the production performance of poultry and bring serious economic losses to the poultry industry. More importantly, this subtype of AIV can pass through host barriers and infect mammals including humans, and it may also become a pandemic in humans, posing a serious threat to human health. At present, t...

Claims

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Application Information

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IPC IPC(8): A61K39/235A61K39/145A61K39/15A61P31/16A61P31/20A61P31/14C12N5/09C12N7/00
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/70C12N5/0693C12N7/00C12N2710/10234C12N2710/10251C12N2720/12034C12N2720/12051C12N2760/16134C12N2760/16151
Inventor 张毓金严悌昆谢秉超敖艳华
Owner 广州博恒生物科技有限公司
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