Method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing bioreactor

A bioreactor, porcine encephalitis technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unreported production methods of porcine encephalitis vaccine, unstable product quality, and intensive manual operations and other issues to achieve the effect of saving manpower, small differences between batches, and reducing costs

Inactive Publication Date: 2011-05-04
WUHAN CHOPPER BIOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The above documents all relate to the preparation method of JE vaccine for humans, but the production process is intensive in manual operation...

Method used

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  • Method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing bioreactor
  • Method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing bioreactor
  • Method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing bioreactor

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0023] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0024] Porcine encephalitis virus: HW1 strain;

[0025] Cell growth medium: M199 (Beijing Qingda Tianyi Biotechnology Co., Ltd.) containing 8% calf serum by volume;

[0026] Virus maintenance solution: M199 (Beijing Qingda Tianyi Biotechnology Co., Ltd.) containing 1% calf serum by volume;

[0027] Cell culture: In 14L bioreactors, add Cytodex1 at a concentration of 10g / L. After hydration, wash with pH 7.2 phosphate buffered saline PBS twice, sterilize, add cell growth medium to balance, and inoculate Vero The cells were cultured; the parameters of the culture method were: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm; samples were taken regularly every day to observe the growth of the cells, and the cells were counted to determine the consumption of glucose. Density up to 1.5×10...

Embodiment 2

[0032] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0033] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0034] Porcine encephalitis virus: HW1 strain;

[0035] Cell growth medium: M199 (Beijing Qingda Tianyi Biotechnology Co., Ltd.) containing 8% calf serum by volume;

[0036] Virus maintenance solution: M199 (Beijing Qingda Tianyi Biotechnology Co., Ltd.) containing 1% calf serum by volume;

[0037] Cell culture: In 14L bioreactors, add Cytodex1 at a concentration of 10g / L, after hydration, wash with pH 7.2 phosphate buffer saline PBS twice, sterilize, add cell growth solution to balance, and inoculate BHK -21 cells were cultured; the parameters of the culture method were: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm; samples were taken regularly every day to observe the growth of the cells, and the cells were counted to determine the consumption of glucose. The density of the ce...

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Abstract

The invention provides a method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing a bioreactor. The method comprises the following steps: (1) sterilizing a microcarrier and the bioreactor, then inoculating cells for vaccine preparation for culturing, inoculating JE viruses when dense monolayers are formed on the cells on the microcarrier, and continuing culturing to reproduce the viruses; (2) stopping culturing and harvesting virus suspension when cytopathy reaches more than 80%; (3) carrying out ultrafilter concentration as well as virus inactivation on the harvested virus suspension; and (4) purifying the inactivated viruses by adopting column chromatography to prepare inactivated vaccines. In the method, the technology of using the microcarrier in the bioreactor for culture is adopted to carry out high density culture of the cells to produce the porcine JE vaccines. Compared with the traditional spinner bottle production method, the method has the following advantages: the automatic control degree is high, so production can be monitored in real time; the labour is saved, thus reducing the cost; the land for production is few, thus being easy to enlarge the scale of production; and the produced viruses have high titer, so the batch-to-batch variation is small, the product quality is stable and the side reaction is low.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for industrially producing porcine Japanese encephalitis vaccine using a bioreactor. Background technique [0002] Japanese Encephalitis (JE), referred to as JE, is a mosquito-borne viral disease that is common to humans and causes reproductive disorders in pigs. Since JEV usually circulates between mosquitoes-pigs-mosquitoes and other animals, pigs are considered to be the most important natural host of JEV, which brings losses to the pig industry and a potential threat to human public health. The current use of vaccination is still an effective means of preventing the disease. [0003] The current large-scale application of porcine JE vaccines include mouse brain inactivated vaccines from Zhongshan strain or Beijing strain virus; cell culture inactivated vaccines from Beijing P-3 strain virus; cell culture live attenuated vaccines from SA14-14- ...

Claims

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Application Information

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IPC IPC(8): A61K39/12C12N7/00A61P31/14C12R1/93
CPCY02A50/30
Inventor 刘汉平秦红刚朱薇温文生漆世华李伟张萍靖志强
Owner WUHAN CHOPPER BIOLOGY
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