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Method for preparing swine fever live vaccines

A technology of swine fever live vaccine and silicone tube, which is applied in the direction of pharmaceutical formula, medical preparations containing active ingredients, recovery/purification, etc., to achieve the effects of improving sensitivity, ensuring titer, and uniform state

Inactive Publication Date: 2013-06-19
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the application of rapid perfusion bioreactor to produce live vaccine against swine fever

Method used

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  • Method for preparing swine fever live vaccines
  • Method for preparing swine fever live vaccines
  • Method for preparing swine fever live vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Cloning, purification and screening of cells for seedling production by limiting dilution method (porcine testis cells ST)

[0039] Porcine testicular cells ST were purchased from China Veterinary Drug Administration;

[0040] The classical swine fever virus vaccine strain is a rabbit-like attenuated strain of classical swine fever virus (strain C), which was purchased from China Veterinary Drug Control Institute;

[0041] The formula of the cell growth solution is 90% high-sugar DMEM solution by volume, 10% newborn bovine serum, and the pH value is adjusted to 7.4;

[0042] The cell maintenance solution is 96% high-sugar DMEM solution by volume, 4% newborn bovine serum, and the pH value is adjusted to 7.4;

[0043] A. Preparation of Monoclonal Cell Lines

[0044] a. Preparation of cell suspension

[0045] Porcine testicular cells were cultured in cell flasks, and when the cells were covered with a dense monolayer and the outline was clear, they were digest...

Embodiment 2

[0065] Embodiment 2 AP20C type torrent perfusion bioreactor prepares swine fever live vaccine

[0066] In the AP20C type torrent perfusion bioreactor used in this example, the perfusion bag 9 has a volume of 4.5 L, the mixer 10 has a volume of 8 L, the torrent tank 11 has a volume of 15 L, and the perfusion bag 9 contains 200 g of polyester fiber paper carrier 8; Theoretical effective culture volume is 18L.

[0067] The cells used for making seedlings are the same as the pig testicular cells (ST) preserved by the strains in Example 1;

[0068] The seed poison is the same as the attenuated strain (C strain) of swine fever rabbitization in Example 1, and a certain batch of seed poison titer is 5×10 5 RID / ml;

[0069] The cell growth solution is 92% high-sugar DMEM solution by volume, 8% newborn bovine serum, and the pH value is adjusted to 7.3;

[0070] The cell maintenance solution is 97% high-sugar DMEM solution by volume, 3% newborn bovine serum, and the pH value is adjust...

Embodiment 3

[0102] Example 3 AP200 Type Torrent Perfusion Bioreactor Preparation of Live Swine Fever Vaccine

[0103] The torrent perfusion bioreactors used in this example include AP20C type and AP200 type.

[0104] The AP20C type torrent perfusion bioreactor is the same as in Example 2.

[0105] The AP200 perfusion bag 9 has a volume of 45L, the mixer 10 has a volume of 80L, the surge tank 11 has a volume of 150L, ​​and the perfusion bag 9 contains 2000g of polyester fiber paper carrier 8; the theoretical effective culture volume is 180L.

[0106] Seedlings are all the same as in Example 2 with cells and seed poisons.

[0107] A. to prepare

[0108] Two torrent perfusion bioreactors, AP20C and AP200, were selected to prepare for cell culture.

[0109] B, preparation work, concrete steps are as follows:

[0110] a. Cultivation of cells for seedling production

[0111] According to the method of Example 2, cultivate the cells for seedling production in the AP20C bioreactor for 96 hou...

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Abstract

The invention relates to a method for preparing swine fever live vaccines, and belongs to the field of biotechnology. The method for preparing the swine fever live vaccines includes the following steps: (1) cell inoculation, (2) culturing of cells used for vaccine preparation, (3) virus inoculation, (4) virus culture and harvest, and (5) vaccine preparation. Cells used for vaccine preparation are screened, adaptability between the cells and virus is strengthened, a riptide perfusion type bioreactor culture system is used for improving multiplication titer and harvest yield of the virus, and a whole production process does not involve other biosafety and public health problems and is suitable for large scale production.

Description

technical field [0001] The invention relates to a method for preparing swine fever live vaccine by using a torrent perfusion bioreactor, belonging to the field of biotechnology Background technique [0002] Classical Swine Fever (CSF), commonly known as "rotten intestinal fever", is called hog cholera in the United States, and classical swine fever in some parts of Europe. The disease is an acute febrile systemic septic infectious disease caused by classical swine fever virus, which can occur in pigs of all ages, and the course of the disease varies from acute to chronic. It is popular throughout the year, highly contagious, and has high morbidity and mortality. Once it occurs, it will be devastating, seriously threatening the development of the pig industry and affecting people's lives. Among swine diseases, swine fever is the most harmful and most important disease. This disease is listed as one of the 16 infectious diseases in category A that must be inspected by the Int...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/187A61P31/14C12N7/02
Inventor 李倬刘涛郑朝朝郁宏伟李建丽杨保收梁武吕茂杰邱贞娜柳珊
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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