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Method for preparing porcine circovirus 2-type inactivated vaccine

A technology of porcine circovirus and inactivated vaccine, which is applied in the directions of antiviral agents, viral antigen components, recovery/purification, etc., can solve problems such as unreported, and achieve the effect of speeding up gas circulation, increasing production, and saving manpower

Active Publication Date: 2015-02-11
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report on the production of porcine circovirus type 2 inactivated vaccine using the rapid perfusion bioreactor

Method used

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  • Method for preparing porcine circovirus 2-type inactivated vaccine
  • Method for preparing porcine circovirus 2-type inactivated vaccine

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 Limiting dilution method for cloning, purifying and screening cells for seedling production

[0039] Pig kidney cells PK15 were purchased from China Veterinary Drug Administration.

[0040] The porcine circovirus vaccine strain is porcine circovirus type 2 ZJ / C strain, which was preserved in the China Center for Type Culture Collection on November 14, 2012. The name of the culture: porcine circovirus type 2 ZJ / C strain ( Porcine cirocovirus, PCV ), the deposit number is CCTCC NO: V201251, and the deposit address is: Wuhan University, China: the virus is relatively common, and the description of its characteristics is omitted.

[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 7.5;

[0043] A. Preparation of Monoclonal ...

Embodiment 2

[0065] Example 2 Preparation of Porcine Circovirus Type 2 Inactivated Vaccine in AP20C Type Torrent Perfusion Bioreactor

[0066] In the AP20C torrent perfusion bioreactor used in this example, the perfusion bag 6 has a volume of 8 L (both perfusion bags have a volume of 4 L), the torrent tank 5 has a volume of 13 L, and the perfusion bag 6 contains 300 g of polyester fiber paper Tablet carrier 8 (two perfusion bags each contain 150g carrier); the theoretical effective culture volume is 21L.

[0067] The pig kidney cell PK15 that cell is preserved with strain in embodiment 1;

[0068] Seed virus is with porcine circovirus type 2 ZJ / C strain in embodiment 1, and a certain batch of seed poison titer is 10 7.5 TCID 50 / ml.

[0069] The formula of the cell growth solution is 92% high-sugar DMEM solution by volume, 8% newborn calf serum, and the pH value is adjusted to 7.4;

[0070] The cell maintenance solution is 98% high-sugar DMEM solution by volume, 2% newborn bovine serum...

Embodiment 3

[0099] Example 3 Preparation of porcine circovirus type 2 inactivated vaccine in AP200 type torrent perfusion bioreactor

[0100] The torrent perfusion bioreactors used in this example include AP20C type and AP200 type.

[0101] The AP20C type torrent perfusion bioreactor is the same as in Example 2.

[0102] The AP200 perfusion bag 6 has a volume of 80L (the volume of the two perfusion bags is 40L), the surge tank 5 has a volume of 130L, and the perfusion bag 6 contains 3000g of polyester fiber paper carrier 8 (each of the two perfusion bags contains 1500g of carrier); Theoretical effective culture volume is 210L.

[0103] Seedlings are all the same as in Example 2 with cells and seed poisons.

[0104] A. Preparation:

[0105] Two torrent perfusion bioreactors, AP20C and AP200, were selected to prepare for cell culture.

[0106] B, preparation work, concrete steps are as follows:

[0107] a. Cultivation of cells for seedling production

[0108] According to the method o...

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Abstract

The invention discloses a method for preparing a porcine circovirus 2-type inactivated vaccine, and belongs to the technical field of organisms. The method comprises the following steps of: a, cultivation of cells for seeding; b, virus inoculation and cultivation; c, virus liquid harvesting; d, virus liquid inactivation; and e, vaccine preparation. By adopting the method, the seeding cells are screened; the matching property of the cells and the virus is enhanced; the multiplication titer and the harvesting yield of the virus are improved by adopting a torrent perfusion-type bioreactor culture system; the immune effect is improved by improvement of an emulsifying process; the entire production process is not involved with other problems of biosafety and public health; and the method is suitable for mass production.

Description

technical field [0001] The invention relates to a method for preparing porcine circovirus type 2 inactivated vaccine by using a torrent perfusion bioreactor, and belongs to the field of biotechnology. Background technique [0002] Porcine circovirus type 2 (PCV2) is the main pathogen of weaned piglet multisystemic wasting syndrome (Postweaning multisystemic wasting syndrome, PMWS). Associated with Nephrotic Syndrome and Porcine Respiratory Disease Syndrome, causing huge economic losses to the swine industry. [0003] At present, the production of porcine circovirus type 2 inactivated vaccine in China is mainly realized by the method of cell spinner bottle culture. The semi-finished product produced by traditional technology has low antigen titer, cumbersome operation, high labor intensity and low efficiency. In recent years, with the development of the veterinary vaccine industry, bioreactors have been widely concerned, and the use of bioreactors to replace the traditional ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61P31/20C12N7/02
Inventor 梁武吕蔚徐倩倩刘涛郑朝朝郁宏伟杨保收李亚杰柳珊赵玉龙朱秀同李建丽邱贞娜
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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