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Method for cultivating attenuated strain pilio inactivated vaccine

A poliomyelitis and culture method technology, which is applied to the culture field of poliomyelitis inactivated vaccine, can solve the problem that cell yield and virus yield cannot meet large-scale production, and achieves good safety, immunogenicity and stability. , the effect of reducing the cultivation space

Active Publication Date: 2007-01-31
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, according to the experience of producing IPV with wild strains, the production of IPV requires a larger amount of antigen than OPV, and the amount of virus used in a single dose is 100 times larger than that of OPV. Large-scale production of attenuated poliovirus inactivated vaccine needs, therefore, it is necessary to develop attenuated polio virus culture technology on microcarriers to meet the needs of children in our country and eradicate polio

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Tertiary amplification cell culture:

[0025] a. First, the cell seeds are centrifugally cleaned, and inoculated into a 7-liter fermenter at a ratio of minimum requirement culture medium (MEM):Vero cell seeds=500:1, and the culture temperature is controlled at 37°C and pH7.2 , dissolved oxygen 50%, and perfusion began on the third day of culture, and the perfusion flow was finally increased from 1 liter / day to 5 liters / day. After 7 days of culture, the cell proliferation reached 2.13×10 6 cells / ml, end the primary culture, and perform trypsinization;

[0026] b. The cell suspension digested with trypsin in step a is inoculated in a 75-liter fermenter at a ratio of minimum requirement culture medium (MEM): Vero cell seed=500: 1, and the culture temperature is controlled at 37°C and pH7. .2, 50% dissolved oxygen, under the condition of 100 liters of recirculation medium (MEM) volume, begin to carry out recirculation culture on the third day, cultivate 7 days, cell den...

Embodiment 2

[0030] 1. Tertiary amplification cell culture:

[0031] a. First, the cell seeds are centrifugally cleaned, and inoculated into a 10 liter fermenter for cell culture according to the ratio of minimum requirement culture medium (MEM): Vero cell seeds=1000: 1, and the culture temperature is controlled at 35° C. and pH 7.0 , dissolved oxygen 30%, and perfusion began on the third day of culture, and the perfusion flow was finally increased from 1 liter / day to 5 liters / day. After 5 days of culture, the cell proliferation reached 2.93×10 6 cells / ml, end the primary culture, and perform trypsinization;

[0032] b. The cell suspension digested with trypsin in step a is inoculated in an 85-liter fermenter at a ratio of minimum requirement culture medium (MEM): Vero cell seed=1000: 1, and the culture temperature is controlled at 35°C and pH7. .0, dissolved oxygen 40%, under the condition of 100 liters of recirculation medium (MEM) volume, begin to carry out recirculation culture on the...

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Abstract

The present invention is cultivation process of deactivated vaccine of attenuated polio strain. The cultivation process is completed with Vero cell and attenuated polio strain virus inside a fermentation tank. Through three stage amplification of cultivation scale, cell cultivating area equivalent to that of 21000 culturing glass bottle of 1 liter volume and cell output equivalent to that of 21000 culturing glass bottle of 1 liter volume may be obtained. The present invention can meet the requirement of large scale preparation of cell and virus liquid. The virus liquid of the present invention may be used to produce vaccine with stable quality, high safety and excellent immunogenicity.

Description

technical field [0001] The invention relates to a method for cultivating an inactivated polio vaccine, belonging to the field of medical biotechnology. Background technique [0002] Poliomyelitis is an acute infectious disease caused by poliovirus types I, II, and III, which is widely spread and extremely harmful. The clinical symptoms cause flaccid paralysis of muscles, especially limbs, which mostly occurs in children, so it is also called "poliomyelitis". Before the vaccine came out, the disease was prevalent all over the world, and the incidence rate in my country was 3.18 / 100,000. In the mid-to-late 1950s, two scientists in the United States, Dr. Salk and Dr. Sabin, successively successfully developed the inactivated polio vaccine (IPV) and the oral polio vaccine (OPV) to prevent and eliminate polio. Yan provides a powerful weapon. Practice has proved that both vaccines are safe and effective. Since 1960, my country began to produce OPV for children all over the cou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/13A61P31/14C12N7/00
CPCY02A50/30
Inventor 褚嘉祐姜述德廖国阳孙明波李卫东张丽旌谢忠平俞泳霆
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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