Method for producing pseudorabies attenuated vaccine by using bioreactor and pseudorabies attenuated vaccine product

A bioreactor and pseudorabies technology, which is applied in the field of veterinary biological products, can solve the problems of inability to achieve culture conditions, low density of cultured cells, and inability to ensure the best conditions for cell culture, achieve uniform and stable product quality, and improve cell culture. Growth density, the effect of good immune protection

Active Publication Date: 2010-04-21
广东永顺生物制药股份有限公司
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Problems solved by technology

[0003] The current production of live pseudorabies vaccine is mainly the traditional culture method of spinner bottle cell culture. This culture method is to cultivate cells to adhere to the wall and grow on the inner wall of the glass culture bottle. Although its operation technology requirements are low and the technology is mature and stable; The culture technology has been used for decades, and it is not suitable for the current large-scale animal vaccine production. The density is small, the cultured virus titer is low, and the immune effect of animals is poor; (2) spinner bottle cell culture has many cell fragments and miscellaneous proteins, and there are many impurities in the vaccine, which leads to large side reactions of immunized animals; (3) spinner bottle cell culture , the difference between cell batches is large, and the product quality is not uniform and unstable; (4) spinner bottle cell culture has low production efficiency and high production cost

Method used

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Embodiment Construction

[0021] In order to make the present invention easier to understand, the present invention will be further described below with reference to specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention, and the specific experimental methods not mentioned in the following examples are usually carried out according to conventional experimental methods.

[0022] Using bioreactor large-scale cell culture technology to produce pseudorabies live vaccine production technology and method, including the following steps:

[0023] (1) Select bioreactor: TideCell fixed bed microcarrier bioreactor.

[0024] (2) Selection of cell lines as cells for seedling preparation: In this example, a porcine testis (ST) cell line was selected as cells for seedling preparation. In the other two examples, porcine kidney (PK15) cell line and porcine kidney (IBRS-2) cell line were selected, respective...

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Abstract

The invention provides a method for producing a pseudorabies attenuated vaccine by using a bioreactor and a pseudorabies attenuated vaccine product. After being sterilized, the bioreactor and a micro carrier are inoculated with cells for producing the vaccine, and a cell growth medium is added for culture. A maintenance medium containing attenuated strains of pseudorabies viruses are inoculated into the bioreactor to continue culturing the cells. 2 to 3 days after virus inoculation, cell culture virus liquid is obtained and added with a stabilizer and antibiotics, and the cell culture virus liquid is refrigerated and dried under vacuum to obtain the pseudorabies attenuated vaccine. In the method, the cell density and virus concentration are improved greatly, the titer of the vaccine is improved, the side reactions, labor intensity and product cost are reduced, the monitoring performance of vaccine production is improved and uniform and stable product quality is guaranteed. The pseudorabies attenuated vaccine produced by the method has high safety, immune efficacy and good immune and protective effect against the attack by the virulent pseudorabies viruses.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to a method for producing a pseudorabies live vaccine using a bioreactor and a product thereof. Background technique [0002] Pseudorabies is an acute infectious disease caused by pseudorabies virus (PRV) in various animals. The infectious disease spreads across the world's major pig-raising countries. Immunization is the main strategy to prevent pseudorabies. [0003] At present, the production of live pseudorabies vaccine is mainly the traditional culture method of rolling bottle cell culture, which is that the cultured cells grow adherently on the inner wall of the glass culture bottle. The culture technology has been used for decades, and it is not suitable for the current large-scale animal vaccine production. The density is low, the titer of cultured virus is low, and the immune effect of animals is poor; (2) The cell culture in a spinner flask has a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245A61P31/20C12N7/00C12R1/93
Inventor 张毓金吴文福林旭野岑小清任向阳游启有刘秋燕陈文胜廖润科刘法连
Owner 广东永顺生物制药股份有限公司
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