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Preparation method of varicella-zoster virus vaccine

A technology for the preparation of herpes zoster virus and vaccine, applied in the field of biomedicine, can solve the problems of poor effect of washing to remove bovine serum albumin, high risk of cell collection and washing aseptic operation, improve efficiency and quality standards, and avoid cell fragmentation , the effect of improving the efficiency of cell collection

Inactive Publication Date: 2020-08-07
江苏金迪克生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a method for preparing a varicella-zoster virus vaccine, so as to solve the problem of high risk of cell collection and aseptic operation in washing and removal of cattle in the production process of varicella-zoster virus vaccine in the prior art. The problem with poor efficacy of serum albumin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Cell resuscitation: MRC-5 cells from ATCC’s working cell bank were resuscitated and passaged. The cell growth medium was M199-801 culture medium containing 10% newborn bovine serum. The cell tube was thawed, and the thawed cell suspension was aseptically inhaled Add growth medium to a 10mL centrifuge tube, and centrifuge at 1000rpm for 5 minutes; discard the supernatant, add cell growth medium to resuspend the cells, transfer to a T25 cell flask, add culture medium, and cultivate at constant temperature for 3 days to obtain the resuscitated cell fluid .

[0023] Cell subculture: First, absorb and discard the liquid in the bottle containing the resuscitated cell liquid, add phosphate buffer solution to wash the cell surface, then add EDTA-Trypsin digestion solution until the cells are covered, and place the cell bottle flat for 1 minute for digestion. When there is a gap in the naked eye, discard the digestive juice, add the cell culture medium, blow and blow the cell su...

Embodiment 2

[0028] Cell resuscitation: MRC-5 cells from ATCC’s working cell bank were resuscitated and passaged. The cell growth medium was M199-801 culture medium containing 15% newborn bovine serum. The cell tube was thawed, and the thawed cell suspension was aseptically inhaled Add growth medium to a 10mL centrifuge tube, and centrifuge at 1000rpm for 5 minutes; discard the supernatant, add cell growth medium to resuspend the cells, transfer to a T25 cell flask, add culture medium, and cultivate at constant temperature for 3 days to obtain the resuscitated cell fluid .

[0029] Cell subculture: First, discard the liquid in the bottle containing the resuscitated cell liquid, add phosphate buffer solution to wash the cell surface, then add EDTA-Trypsin digestion solution until the cells are covered, and place the cell bottle flat for 2 minutes for digestion. When gaps appear on the naked eye, discard the digestive juice, add cell culture medium, blow and blow the cell surface to disperse...

Embodiment 3

[0034] Cell resuscitation: MRC-5 cells from ATCC’s working cell bank were resuscitated and passaged. The cell growth medium was M199-801 culture medium containing 20% ​​newborn bovine serum. The cell tube was thawed, and the thawed cell suspension was aseptically inhaled Add growth medium to a 10mL centrifuge tube, and centrifuge at 1000rpm for 5 minutes; discard the supernatant, add cell growth medium to resuspend the cells, transfer to a T25 cell flask, add culture medium, and cultivate at constant temperature for 4 days to obtain the resuscitated cell fluid .

[0035] Cell subculture: First, discard the liquid in the bottle containing the resuscitated cell liquid, add phosphate buffer solution to wash the cell surface, then add EDTA-Trypsin digestion solution until the cells are covered, and place the cell bottle flat for 3 minutes for digestion. When there is a gap in the naked eye, discard the digestive juice, add the cell culture medium, blow and blow the cell surface to...

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Abstract

The invention discloses a preparation method of a varicella-zoster virus vaccine. The preparation method comprises the following specific preparation steps: carrying out centrifugal separation on harvested cell sap, a UniFuge disposable barrel-shaped bowl body centrifugal machine is adopted for centrifugal separation, in addition, the method further comprises the procedures of cell recovery, cellpassage, virus culture, cell harvesting and centrifugal washing. According to the invention, the problems of high cell collection and washing sterile operation risk and poor bovine serum albumin washing and removal effect in the varicella-zoster virus vaccine production process in the prior art can be solved.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for preparing a varicella-zoster virus vaccine. Background technique [0002] The live attenuated varicella vaccine is a freeze-dried virus product obtained by culturing and propagating varicella virus OKA strain in MRC-5 diploid cells. It is suitable for active immunization against chickenpox for healthy children over 12 months old, adolescents and adults, high-risk groups and their close contacts. The live attenuated herpes zoster vaccine is basically similar to the attenuated live varicella vaccine, and its concentration is about 10 times that of the varicella vaccine. [0003] The Food and Drug Administration strongly encourages and guides manufacturers to improve and ensure the quality of vaccines through disposable production technology. At present, the existing technology of varicella-zoster virus vaccine is to cultivate human diploid cells in the cell factory and Cor...

Claims

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Application Information

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IPC IPC(8): C12N7/02A61K39/25A61P31/22C12R1/93
CPCA61K39/12A61P31/22C12N7/00C12N2710/16734C12N2710/16751
Inventor 杨骏宇余军望朔朱实惠杨文彬唐阳赵永强李凡
Owner 江苏金迪克生物技术股份有限公司
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