Method for preparing porcine parvovirus inactivated vaccines

A technology of parvoviruses and inactivated vaccines, applied in antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc., can solve problems such as unreported methods, improve immune effect, increase adherent area, The effect of reducing usage

Active Publication Date: 2014-07-23
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But, there is no report on the method of producing porcine parvovirus disease inactivated vaccine using the rapid perfusion bioreactor

Method used

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  • Method for preparing porcine parvovirus inactivated vaccines
  • Method for preparing porcine parvovirus inactivated vaccines
  • Method for preparing porcine parvovirus inactivated vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043]Example 1 Cloning, purification and screening of cells for seedling production by limiting dilution method (pig kidney cell IBRS-2)

[0044] Pig kidney cells IBRS-2 were purchased from China Veterinary Drug Administration.

[0045] The porcine parvovirus vaccine strain is porcine parvovirus RP2 strain, which was purified and screened by plaque method, and was preserved in the China Type Culture Collection Center on November 14, 2012. The culture name: Porcine parvovirus RP2 strain (Porcine parvovirus, PPV ), the deposit number is CCTCC NO: V201250 and the deposit address is Wuhan University, China. The virus is a common virus in the field, so the description of its characteristics is omitted.

[0046] 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;

[0047] The cell maintenance solution is 96% high-sugar DMEM solution by volume, 4% newborn bovine serum, and the pH value is...

Embodiment 2

[0075] Example 2 Preparation of porcine parvovirus inactivated vaccine in AP20C type torrent perfusion bioreactor

[0076] 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.

[0077] Cell is the pig kidney cell IBRS-2 preserved with strain in embodiment 1;

[0078] The seed virus is the same as the porcine parvovirus RP2 strain in embodiment 1;

[0079] 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;

[0080] The cell maintenance solution is 98% high-sugar DMEM solution by volume, 2% newborn bovine serum, and the pH value is adjusted to 7.5;

[0081] Na-DMEM dry powder m...

Embodiment 3

[0109] Example 3 Preparation of porcine parvovirus inactivated vaccine by AP200 type torrent perfusion bioreactor

[0110] The torrent perfusion bioreactors used in this example include AP20C type and AP200 type.

[0111] The AP20C type torrent perfusion bioreactor is the same as in Example 2.

[0112] 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.

[0113] Seedlings are all the same as in Example 2 with cells and seed poisons.

[0114] A. Preparation:

[0115] Two torrent perfusion bioreactors, AP20C and AP200, were selected to prepare for cell culture.

[0116] B, preparation work, concrete steps are as follows:

[0117] a. Cultivation of cells for seedling production

[0118] According to the method of Examp...

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Abstract

The invention relates to a method for preparing porcine parvovirus inactivated vaccines, and belongs to the field of biotechnology. The method for preparing the porcine parvovirus inactivated vaccines includes the following steps: (1) culturing of cells used for vaccine preparation, (2) virus inoculation and culture, (3) virus liquid harvest, (4) virus liquid inactivation, and (5) vaccine preparation. Seed virus used for vaccine preparation and cells are screened, matching degree between the virus and the cells is strengthened, a riptide perfusion type bioreactor culture system is used for improving multiplication titer and harvest yield of the virus, immune effects are improved through the improvement of an emulsification process 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 an inactivated porcine parvovirus vaccine by using a torrent perfusion bioreactor, which belongs to the field of biotechnology. Background technique [0002] Porcine parvovirus (Porcineparvo viurs, PPV) is caused by porcine parvovirus infection, which can cause reproductive disorders in pigs, mainly manifested as abortion, stillbirth, infection and death of fetuses and embryos in pregnant sows, and the mother usually does not No clinical symptoms. In pig herds all over the world, the virus is ubiquitous, generally endemic or sporadic in most pig farms, and it is difficult to eradicate and cause huge economic losses to the pig industry. There is no effective treatment for PPV infection, so the prevention and immunization of the disease is even more important. [0003] It is recognized that the use of vaccines is an effective way to prevent porcine parvovirus and improve the reproductive rate of sows. At...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/23A61P31/20C12N7/02
Inventor 郑朝朝李晓艳刘涛刘超郁宏伟李建丽杨保收梁武柳珊
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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