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Method for industrially producing swine fever live vaccine by using bioreactor

A bioreactor and swine fever live vaccine technology, applied in the field of veterinary biological products, to achieve the effects of easy scale expansion, cost reduction, and stable product quality

Active Publication Date: 2013-07-03
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the use of stirred bioreactors and the inverted tank operation technology of step-by-step amplification for high-density culture of cells to prepare vaccines.

Method used

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  • Method for industrially producing swine fever live vaccine by using bioreactor
  • Method for industrially producing swine fever live vaccine by using bioreactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0022] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0023] Classical swine fever virus: attenuated strain of classical swine fever rabbit;

[0024] Cell growth medium: DMEM containing 8% calf serum (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0025] Virus maintenance solution: DMEM containing 1% calf serum by volume; (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0026] Cell culture: In 14L bioreactors, add Cytodex-1 microcarriers at a concentration of 10g / L. After hydration, wash with pH 7.2 phosphate buffer solution PBS several times, and add cell growth solution to balance after sterilization. , Inoculation of SK6 cells for culture. The parameters of the culture method are: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm; samples are taken regularly every day to observe the growth of the cells, count the cells, and meas...

Embodiment 2

[0031] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0032] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0033] Classical swine fever virus: attenuated strain of classical swine fever rabbit;

[0034] Cell growth medium: DMEM / F12 (Hyclone) containing 10% calf serum by volume;

[0035] Virus maintenance solution: DMEM / F12 (Hyclone) containing 2% calf serum by volume;

[0036] Cell culture: In 14L bioreactors, add Cytodex-1 microcarriers at a concentration of 10g / L. After hydration, wash with pH 7.2 phosphate buffer solution PBS several times, and add cell growth solution to balance after sterilization. 1. Inoculate PK-15 cells for culture. Take regular samples every day to observe the cell growth, count the cells, and measure the consumption of glucose. When the cell density reaches 1.5×10 6 perml, start perfusion, the rate of perfusion is 0.5-5 working volumes per day according to the density of cells and th...

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Abstract

The invention relates to a method for industrially producing a swine fever live vaccine by using a bioreactor. The method comprises the following steps of: (1) sterilizing a microcarrier and the bioreactor, adding cell growth solution, inoculating cells for vaccine production, culturing the cells, and after the cells on the microcarrier form a compact monolayer, discarding the cell growth solution, adding virus maintenance solution and inoculating swine fever virus, and continuously culturing to allow the virus to reproduce; and (2) after the virus is inoculated for 18 hours, continuously obtaining virus culture solution, storing the virus culture solution at the temperature of below -15 DEG C, mixing the virus culture solution, and adding a cryoprotectant to prepare the swine fever live vaccine. The method has the advantages of high controllability of process parameters, high cell culture density, high virus titer, high vaccine safety, stable and reliable quality, high production efficiency and the like.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for industrially producing live swine fever vaccines using bioreactors. Background technique [0002] Classical swine fever virus (CSFV) is a highly contagious disease caused by classical swine fever virus (CSFV), which poses a huge threat to the swine industry worldwide. Pigs of different ages, sexes and breeds can be infected, and the mortality rate is as high as 80-90%. In my country, the prevalence of CSF presents the coexistence of typical CSF and atypical CSF, the coexistence of persistent infection and recessive infection, and the coexistence of immune tolerance and virus-carrying syndrome. The new epidemic form of swine fever has brought new challenges to my country's pig industry, and immunization is still an important measure to prevent and control swine fever. [0003] The current production of live swine fever vaccine mainly adopts th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/187C12N7/00A61P31/14C12R1/93
Inventor 漆世华刘汉平秦红刚肖敏温文生段振华王威张萍
Owner WUHAN CHOPPER BIOLOGY
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