Large-scale production method of porcine transmissible gastroenteritis virus

A production method and infectious technology, applied in biochemical equipment and methods, viruses/bacteriophages, microorganisms, etc., can solve problems such as low virus titer, low virus yield, and large batch-to-batch differences, and achieve low cell inoculum volume , high virus infection efficiency and small batch-to-batch difference

Active Publication Date: 2013-05-08
PU LIKE BIO ENG
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Problems solved by technology

[0005] In view of this, the main purpose of the present invention is to provide a large-scale production method of porcine transmissible gastroenteritis virus with reasonable technology, high virus yield and titer, which can solve the problem of traditional rotary bottle culture mode in virus production. The existing shortcomings of low virus yield, low virus titer, and large batch-to-batch difference can also realize continuous production of virus, and achieve high-titer multiple harvest of virus liquid by optimizing parameters, which greatly improves the yield of virus

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  • Large-scale production method of porcine transmissible gastroenteritis virus

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Embodiment 1

[0044] Example 1 Large-scale production of porcine transmissible gastroenteritis virus by tidal microcarrier suspension bioreactor

[0045] 1. Viruses and cell lines

[0046] The porcine transmissible gastroenteritis virus used to prepare porcine transmissible gastroenteritis (TGE) Huadu attenuated virus (H) vaccine strain (purchased from the Chinese Type Culture Collection Center of Wuhan University, preservation number CCTCC-V200609), after pathogenic Sex test, the strain virus is not pathogenic. The porcine testis (ST) cell line, which has good sensitivity to the virus strain, is used as the cell line for making seedlings, so as to infect and multiply porcine transmissible gastroenteritis virus in large quantities.

[0047] 2. Preparation method

[0048] (1) Inoculate the porcine testis cell line (ST), the cell line used for making seedlings, into a carrier with DMEM liquid medium (including 5% bovine serum, double antibody with a final concentration of 100IU, and a pH ...

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Abstract

The invention provides a large-scale production method of a porcine transmissible gastroenteritis virus. The method comprises the steps that: in a tidal-type bioreactor, cells are inoculated to micro-carriers, such that cell absorption culturing and amplification culturing are carried out; porcine transmissible gastroenteritis virus is inoculated, and virus absorption culturing is carried out; virus proliferation culturing is carried out; and when cell CPE reaches 70% or higher, virus solution is collected. The porcine transmissible gastroenteritis virus large-scale production method provided by the invention shows a trend of animal cell and virus continuous culturing and large-scale production. Compared with a traditional shake flask culturing process, an antigen titer is improved by more than 30 times. Product quality is homogenous, production process is simplified, operation is simple, production efficiency is improved, and production cost is reduced.

Description

technical field [0001] The invention relates to a large-scale production method of porcine transmissible gastroenteritis virus, which belongs to the field of veterinary vaccines. Background technique [0002] Porcine transmissible gastroenteritis virus (TGEV) is a member of the Coronaviridae family and the genus Coronaviridae. TGEV can infect pigs of all ages, especially causing vomiting, severe diarrhea and high mortality in piglets under the age of two weeks. It is a highly contagious and highly contagious intestinal infectious disease of pigs that is widespread in the world, and often occurs repeatedly in pig farms. Occurrence and prevalence bring great harm to the pig industry (Pensaert M B. Disease of Swine. Seventh edition. Iowa State University Press. USA. 1992, 305-338). In the research and clinical discussion of many scholars, it is found that the effect of using drugs on porcine transmissible gastroenteritis is not very clear. The key to controlling the disease is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/02
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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