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Method for industrially producing pseudorabies vaccine by using bioreactor

A bioreactor, pseudorabies technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effects of cost reduction, easy expansion of scale, and small batch-to-batch differences

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

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Problems solved by technology

However, there is no report on the use of stirred bioreactors and the inverted tank operation technology of step-by-step amplification for high-density culture of cells to prepare vaccines.

Method used

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  • Method for industrially producing pseudorabies vaccine by using bioreactor
  • Method for industrially producing pseudorabies vaccine by using bioreactor
  • Method for industrially producing pseudorabies vaccine by using bioreactor

Examples

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Embodiment 1

[0021] Example 1: Bioreactor: 14L and 40L bioreactors from NBS Company in the United States, the parameters of the bioreactor are: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm;

[0022] Microcarrier: Cytodex1, General Electric Healthcare Life Sciences Division;

[0023] Pseudorabies virus: Bartha-K61 strain;

[0024] Cell growth medium: DMEM containing 8% calf serum (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0025] Virus maintenance solution: DMEM containing 1% calf serum by volume (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0026] Cell culture: In 14L bioreactors, add Cytodex-1 at a concentration of 10g / L, after hydration, wash with pH 7.4 phosphate buffered saline PBS several times, sterilize, inoculate ST cells, and culture ; Take regular samples every day to observe the growth of the cells, count the cells, and measure the consumption of glucose. When the density of the cells reaches 1.5×10 6 perml, start perfusion, and the per...

Embodiment 2

[0030] Embodiment 2: Bioreactor: 14L and 40L bioreactors of NBS Company in the United States, the parameters of the bioreactor are: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm;

[0031] Microcarrier: Cytodex1 (General Electric Healthcare Life Sciences);

[0032] Pseudorabies virus: E A virulent strain

[0033] Cell growth medium: DMEM / F12 containing 8% calf serum (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0034] Virus maintenance solution: DMEM / F12 (Beijing Qingda Tianyi Biotechnology Co., Ltd.) containing 1% calf serum by volume;

[0035] Cell culture: In 14L bioreactors, add Cytodex1 at a concentration of 10g / L, after hydration, wash with pH7.4 phosphate buffered saline PBS several times, sterilize, inoculate ST cells, and cultivate; Take samples at regular intervals to observe cell growth, count cells, and measure glucose consumption. When the cell density reaches 1.5×10 6 / ml, start perfusion, and the perfusion speed depends on the d...

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Abstract

The invention relates to a method for industrially producing a pseudorabies vaccine by using a bioreactor. The method comprises the following steps of: (1) sterilizing a microcarrier and the bioreactor, inoculating cells for vaccine production, culturing the cells, and after the cells on the microcarrier form a compact monolayer, inoculating pseudorabies virus, and continuously culturing to allow the virus to reproduce; (2) after more than 80 percent of cells are subjected to a cytopathic effect, stopping culturing, and obtaining virus liquid to prepare the pseudorabies vaccine. In the method, the pseudorabies vaccine is produced by culturing the cells in high density by the bioreactor microcarrier culture technology; and compared with the conventional roller bottle production process, the invention has the advantages that: the method has high automation control degree, saves manpower, reduces cost and is small in production land, the production can be monitored in real time, the scale is easy to expand, the produced virus has high titer and small difference among batches, and the product has stable quality and small side effect.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a method for industrially producing pseudorabies vaccine using a bioreactor. Background technique [0002] Porcine pseudorabies (Pseudorabies, Pr) is an infectious disease caused by porcine herpesvirus type I pseudorabies virus of the family Herpesviridae. my country lists it as a second-class animal disease. Pseudorabies has been prevalent in the world and widely exists in our country. Many large-scale pig farms use pseudorabies vaccine to prevent and control the disease. [0003] At present, the production of pseudorabies vaccine mainly adopts the traditional spinning bottle process, that is, the pseudorabies virus is inoculated into cells that have formed a monolayer, and the virus liquid is harvested at one time after cultivation. However, each spinner bottle is an independent cell culture unit, and the quality of cells, virus yield and ti...

Claims

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Application Information

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IPC IPC(8): A61K39/245C12N7/00A61P31/22C12R1/93
Inventor 秦红刚刘汉平李伟肖敏王桢桢漆世华温文生
Owner WUHAN CHOPPER BIOLOGY
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