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Method for production of porcine epidemic diarrhea virus

A technology for porcine epidemic diarrhea and a production method, which is applied to the production technology field of the rolling bottle culture method, can solve the problems of polluted vaccine quality, easy to be polluted by the environment, and high production cost, and achieves uniform product quality, reduced production cost, and short production cycle. Effect

Active Publication Date: 2011-08-10
成都史纪生物制药有限公司
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Problems solved by technology

The traditional process is labor-intensive, time-consuming, low in efficiency, and high in production cost; it is easy to be polluted by the environment; there are large differences between different production batches or between different bottles of the same production batch; it is difficult to expand production; it is easy to be contaminated by bacteria or other viruses The hidden dangers of vaccine quality, involving biosafety and public health issues

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  • Method for production of porcine epidemic diarrhea virus
  • Method for production of porcine epidemic diarrhea virus

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Embodiment Construction

[0024] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0025] (1) Choose a bioreactor as the means of cultivation: 75L bioreactor.

[0026] (2) Microcarriers are selected as the carrier for cell-attached growth: Microcarrier Cytodex-2

[0027] (3) Cleaning and sterilization methods of microcarriers: 1) Weigh 10 g / L of Cytodex-2 microcarriers and soak in 2L of PBS solution for three hours. 2) Wash 3 times with 2L PBS each time. 3) Add 2L of PBS solution to soak the microcarriers, and steam sterilize at 121° C. for 30 minutes.

[0028] (4) Select VERO cells as cells for making seedlings:

[0029] (5) Passage and culture of cells for seedling production: the above cells were digested and passaged by EDTA-trypsin cell dispersion (Hank's solution containing 0.25% trypsin (specification: 1:250), 0.02% EDTA). 92% DMEM solution, 8% calf serum, 200 IU / ml sodium penicillin and streptomycin sulfate, and t...

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Abstract

The invention discloses a technology for the production of porcine epidemic diarrhea virus by means of the microcarrier culture of VREO cells using a bioreactor, and comprises the technology for the production of different porcine epidemic diarrhea virus strains. The technology comprises the following technical steps: (1) selection of VERO cells as cell line for vaccine; (2) passage and culture of cells for vaccine; (3) propagation of seed culture of the porcine epidemic diarrhea virus; (4) microcarrier suspension culture of the VERO cells in the bioreactor; (5) propagation of porcine epidemic diarrhea virus antigen; and (6) treatment of acquired virus antigen liquid. The production method can remarkably lower production cost and enhance output-input ratio by 5 to 10 times, and has the advantages of short production period, small occupied space, great easiness for enlarging production scale rapidly, little environmental pollution, easy processing, high automation degree, a small number of staff, easy implementation of even and stable quality, obviously lowered production cost and enhanced yield and quality of vaccine.

Description

technical field [0001] The invention relates to a method for producing vaccines using bioreactor microcarrier cell culture technology, which can be used for industrial production of porcine epidemic diarrhea vaccines and replaces the traditional production process of rotary bottle culture method. Background technique [0002] At present, the production of porcine epidemic diarrhea vaccine in China only relies on the cell spinner bottle culture method. The traditional process is labor-intensive, time-consuming, low in efficiency, and high in production cost; it is easy to be polluted by the environment; there are large differences between different production batches or between different bottles of the same production batch; it is difficult to expand production; it is easy to be contaminated by bacteria or other viruses The hidden dangers of vaccine quality caused by this problem involve biosafety and public health issues. Contents of the invention [0003] The purpose of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12R1/91C12N7/02C12R1/93C12N7/00
Inventor 徐宏军胡来根刘玉才
Owner 成都史纪生物制药有限公司
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