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A kind of method for preparing rabies vaccine

A technology of rabies vaccine and torrent perfusion, which is applied in the field of rabies vaccine preparation, can solve problems such as unreported rabies vaccine, and achieve the effects of optimizing cell growth environment, good immunogenicity and ensuring titer.

Active Publication Date: 2015-08-12
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 in the production of rabies vaccine

Method used

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  • A kind of method for preparing rabies vaccine
  • A kind of method for preparing rabies vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Rabies virus rejuvenation

[0040] BHK21 cells, rabies virus Flury-LEP strain, and Paris strain rabies fixed virus were all purchased from China Veterinary Drug Control Institute.

[0041] The formula of the cell growth solution is 92% high-sugar DMEM solution by volume, 8% newborn bovine serum, and the pH value is adjusted to 7.2;

[0042] The cell maintenance solution is 99% high-sugar DMEM solution by volume, 1% newborn bovine serum, and the pH value is adjusted to 7.3;

[0043] a. Rabies virus returns to mouse brain

[0044] Rabies virus Flury-LEP strain seed, with PBS (0.04mol / L, pH value 7.6) 10 times serial dilutions, get 10 -2 , inoculate 11-13g of mice, inoculate into the brain, 0.03mL / monkey, feeding observation, choose to inoculate the mice with typical rabies symptoms for 4-5 days, take the brain tissue aseptically, grind, add serum-free DMEM solution to make 20 % suspension, centrifuge at 6000r / min for 20min, take the supernatant, and store it ...

Embodiment 2

[0059] Example 2 Preparation of Live Rabies Vaccine by AP20C Type Torrent Perfusion Bioreactor

[0060] 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.

[0061] BHK21 cells are the same as in Example 1;

[0062] Seed virus is with the rabies virus Flury-LEP strain that bacterial classification preserves in embodiment 1, and a certain batch of seed poison titer is 10 5.8 LD 50 / 0.03ml.

[0063] 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 7.3;

[0064] The cell maintenance solution is 98% high-sugar DMEM solution by volume, 2% newborn bovine serum, and the pH value is adjusted to 7.4;

[0065] The Na-DMEM dry powder medium ...

Embodiment 3

[0094]Example 3 Preparation of inactivated rabies vaccine by AP200 type torrent perfusion bioreactor

[0095] The torrent perfusion bioreactors used in this example include AP20C type and AP200 type.

[0096] The AP20C type torrent perfusion bioreactor is the same as in Example 2.

[0097] 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.

[0098] Seedlings are all the same as in Example 2 with cells and seed poisons.

[0099] A. to prepare

[0100] Two torrent perfusion bioreactors, AP20C and AP200, were selected to prepare for cell culture.

[0101] B, preparation work, concrete steps are as follows:

[0102] a. Cultivation of cells for seedling production

[0103] According to the method of Example 2, cultivate the cells for seedling production in the AP20C bioreactor for 12...

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Abstract

The invention discloses a method for preparing a rabies vaccine, and belongs to the technical field of organisms. The method comprises the following steps of: a, cell inoculation; b, cultivation of cells for seeding; c, virus vaccination, d, virus cultivating and harvesting; and e, vaccine preparation. By adopting the method, the virus is rejuvenated; the reproductive capacity of 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 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 rabies vaccine by using a torrent perfusion bioreactor, belonging to the field of biotechnology Background technique [0002] Rabies is a zoonotic disease caused by rabies virus that all warm-blooded animals are susceptible to. The disease occurs and is prevalent worldwide. Rabies virus is transmitted through animal bites and scratches. The virus attacks the nerves of animals and humans system, once the disease occurs, the mortality rate is about 100%. my country is an area with a high incidence of rabies in the world, and the death toll of rabies has always occupied the first place in my country's legally notifiable infectious diseases. It is a major epidemic that endangers my country's public health security. [0003] Dogs and cats play an important role in carrying and spreading rabies, and are the main source of human and animal infection with rabies virus. Improving the rabies vaccine immunization ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/245A61K39/205A61P31/14A61P31/22C12M3/00
Inventor 邱贞娜刘涛郑朝朝郁宏伟柳珊韩佳丽杨保收梁武李建丽
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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