A kind of method of producing porcine pseudorabies live vaccine with passage cell line

A technology of passaging cells and cells, which is applied in the field of producing porcine pseudorabies live vaccines by using passaging cell lines, can solve the problems of easy carrying of exogenous viruses, etc., facilitate transportation and storage, have high stability, and overcome low virus titers high effect

Active Publication Date: 2019-09-06
广州渔跃生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, ST cells are derived from pigs and are easy to carry exogenous viruses, which has a great impact on the production of porcine pseudorabies live vaccines

Method used

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  • A kind of method of producing porcine pseudorabies live vaccine with passage cell line

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Effect test

Embodiment 1

[0027] Embodiment 1 A kind of method of producing porcine pseudorabies live vaccine with passage cell line

[0028] The method for producing porcine pseudorabies live vaccine with the passage cell line is divided into the following steps:

[0029] S1. Passaging and culturing of Vero cells for seedling production: Digest the passaging cells with EDTA-trypsin cell dispersion liquid, continue to culture with cell growth medium, and form a monolayer for continued passaging or virus inoculation;

[0030] The cell growth liquid consists of the following components and their contents: 89% of DMEM liquid, 3% of Sapindus saponin and 8% of newborn bovine serum; and the pH value is 7.2;

[0031] S2. Propagation of Vero cell virus species: inoculate the well-grown above-mentioned passaged cell monolayer with the attenuated pseudorabies virus strain, and continue to cultivate with cell maintenance medium; harvest the cell culture venom after 3 days as the virus species for production;

[...

Embodiment 2

[0036] Embodiment 2 A kind of method of producing porcine pseudorabies live vaccine with passage cell line

[0037] The method for producing porcine pseudorabies live vaccine with the passage cell line is divided into the following steps:

[0038] S1. Passaging and culturing of Vero cells for seedling production: Digest the passaging cells with EDTA-trypsin cell dispersion liquid, continue to culture with cell growth medium, and form a monolayer for continued passaging or virus inoculation;

[0039] The cell growth liquid includes the following components and their contents: 90% of DMEM liquid, 1% of Sapindus sapinsin and 9% of newborn bovine serum;

[0040] S2. Propagation of Vero cell virus species: inoculate the well-grown above-mentioned passaged cell monolayer with the attenuated pseudorabies virus strain, and continue to cultivate with cell maintenance medium; harvest the cell culture venom after 3 days as the virus species for production;

[0041] The cell maintenance ...

Embodiment 3

[0045] Embodiment 3 A kind of method of producing porcine pseudorabies live vaccine with passage cell line

[0046] The method for producing porcine pseudorabies live vaccine with the passage cell line is divided into the following steps:

[0047] S1. Passaging and culturing of Vero cells for seedling production: Digest the passaging cells with EDTA-trypsin cell dispersion liquid, continue to culture with cell growth medium, and form a monolayer for continued passaging or virus inoculation;

[0048] The cell growth liquid includes the following components and their contents: 85% of DMEM liquid, 5% of Sapindus saponin and 8.7% of neonatal bovine serum;

[0049] S2. Propagation of Vero cell virus species: inoculate the well-grown above-mentioned passaged cell monolayer with the attenuated pseudorabies virus strain, and continue to cultivate with cell maintenance medium; harvest the cell culture venom after 3 days as the virus species for production;

[0050] The cell maintenance ...

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Abstract

The invention especially relates to a method for producing a live vaccine against pseudorabies in swine from a continuous cell line, belonging to the technical field of biological products for livestock. The continuous cell line used in the invention is Vero cells. The method comprises the following steps: passage and culture of cells for vaccine preparation; propagation of virus seeds of the cells; propagation of virus liquid used for vaccine preparation; and vaccine preparation, split charging and freeze-drying. Porcine pseudorabies viruses obtained by using the method re high in content and titer; and the eventually prepared vaccine has high stability and long shelf life.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a method for producing porcine pseudorabies live vaccine by using subcultured cell lines. Background technique [0002] Porcine pseudorabies is an acute infectious disease caused by porcine pseudorabies virus. The epidemic spreads throughout the world's major pig-growing countries. Immunization is the main measure to prevent porcine pseudorabies. At present, the cells used in the production of porcine pseudorabies live vaccines in my country are chicken embryo fibroblasts and ST cells. Due to the difference in raw materials of each batch of chicken embryos, the difference between batches of chicken embryo fibroblast primary cells is large; the virus titer of porcine pseudorabies virus is not high when cultured with chicken embryo fibroblasts, which brings great benefits to the improvement of vaccine production and efficacy. Difficult, and the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/245A61K47/42A61P31/22C12N5/071C12N7/00
Inventor 张毓金严悌昆黄淑芬
Owner 广州渔跃生物技术有限公司
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