Production method of pseudorabies virus vaccine

Abstract

The invention discloses a production method of a pseudorabies virus vaccine. The production method comprises the following steps: (1) adopting a cell of a continuous cell line, subjecting the cell to digestive passage, and continuously culturing the cell in a cell culture flask by using a cell growth medium; (2) diluting a virus seed into 10 times by using a cell maintenance medium, and inoculating a cell monolayer to obtain an infected cell virus liquid, that is, a virus seed for production; (3) preparing the cell monolayer formed in the step (1) into a cell suspension through digestion, inoculating the cell suspension into a bioreactor, and adding a microcarrier into the bioreactor; (4) performing virus-introduction operation on the cell, wherein the introduced virus seed is the virus seed produced in the step (2); and (5) harvesting liquid in the reactor together with the microcarrier until all of the cells on the microcarrier fall off and a dissolved oxygen value increases obviously, placing the liquid and the microcarrier under conditions of minus 20 DEG C, repeatedly freezing and thawing the liquid and the microcarrier twice, and removing the microcarrier and cell fragments to prepare the pseudorabies virus vaccine. The production method of the pseudorabies virus vaccine is short in production cycle and large in yield.

Description

technical field [0001] The embodiment of the present invention relates to a production method of a pseudorabies virus vaccine, in particular to a method for producing a pseudorabies virus vaccine by using microcarrier cell suspension culture in a bioreactor. Background technique [0002] At present, domestic pseudorabies virus vaccine production only relies on the spinner bottle culture method. The traditional process is labor-intensive, time-consuming, low in efficiency, and high in production cost; it is easy to be polluted by the environment; the difference between different batches of spinner bottle culture is large; it requires a lot of space; it may be contaminated by bacteria or other viruses during the spinner bottle culture operation Increased toxicity may lead to quality problems of the vaccines produced; once contamination occurs during spinner bottle cultivation, due to the large number of spinner bottles, it is difficult to harmlessly dispose of them, which invo...

AI Technical Summary

Problems solved by technology

The traditional process is labor-intensive, time-consuming, low in efficiency, and high in production cost; it is easy to be polluted by the environment; the difference between different batches of spinner bottle culture is large; it requires a lot of space; it may be contaminated by bacteria or other viruses during the spinner bottle culture operation Increased toxicity may lead to quality problems of the vaccines produced; once contamination occurs during spinner bottle cultivation, due to the large number of spinner bottles, it is difficult to harmlessly dispose of them, which involves biosafety and public health issues

In addition, there have been reports on the use of bioreactor microcarriers to cultivate rabies vaccines, but there has been no report on the use of bioreactor microcarriers to produce pseudorabies virus

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