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Method for preparing inactivated rabies vaccine for animal by bioreactor

A bioreactor and inactivated vaccine technology is applied in the field of preparing veterinary rabies inactivated vaccines using bioreactors, which can solve the problems of short cell maintenance time, unfavorable quality improvement, and reduced production costs, achieving quality controllable, The effect of low cost and stable production process

Inactive Publication Date: 2013-03-06
TANGSHAN YIAN BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the use of rabies vaccine varieties, the attenuated vaccine was used before. After a period of use, due to the inherent weakness of the attenuated vaccine, its scope of use is limited
[0005] At present, the production of attenuated rabies vaccines in my country has problems such as intensive manual operations, limited virus strains, short cell maintenance time, low density and activity, which limit the improvement of product output and quality and are not conducive to reducing production costs.

Method used

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  • Method for preparing inactivated rabies vaccine for animal by bioreactor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1 Vero cell subculture expansion culture adapted to rabies virus CTN-1 strain

[0017] Rabies virus CTN-1 strain was inoculated in Vero cells at a ratio of 0.5%, cultured at 34.0±2.0°C, the supernatant was harvested, frozen, and continuously passaged on Vero cells in the same way. The virus titer was determined to reach 10 by intracerebral inoculation in mice. 8 LD 50 / ml above, indicating that the CTN-1 strain virus is sensitive to Vero cells.

Embodiment 2

[0018] Embodiment 2 utilizes bioreactor and Cytodex-1 microcarrier system to carry out Vero cell culture

[0019] The working volume of the bioreactor is 10L-500L, the Cytodex-1 microcarrier system uses 10g / L, and the cell density ranges from 5×10 5 Increase to 5~7×10 6 .

Embodiment 3

[0020] Embodiment 3 Inoculation of rabies virus CTN-1 strain carries out virus infection to Vero cells

[0021] After the CTN-1 strain rabies virus infected Vero cells, according to the glucose concentration, lactate concentration, glutamine, oxygen consumption rate and pH value index in the assay medium (containing 1-5% bovine serum standard MEM medium) The growth of Vero cells is controlled; the virus culture temperature is 34.0±2.0° C., the pH value is 7-8, the dissolved oxygen is 10-100%, and the stirring speed is 10-100 rpm.

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Abstract

The invention provides a method for preparing inactivated rabies vaccine for animal by a bioreactor. The method includes by the aid of the bioreactor and a Cytodex-1 microcarrier system, subjecting high-density Vero cells (or BHK cells) adaptive to rabies virus CTN-1 strain (aG-strain or PV-strain) to virus infection by utilizing the CTN-1 strain (aG-strain or PV-strain) rabies virus with good antigenicity so as to prolong virus harvest time from 6-9 days to 11-15 days, increase virus titer from 105-7LD50 / ml to 108-9LD50 / ml and increase virus content by 10-1000 times; and inactivating harvest virus liquid, adding additives, and then packing to obtain high-quality vaccine products. The prepared vaccine is safe and effective, production process is stable, and production quality is controllable.

Description

technical field [0001] The invention relates to a preparation method of a vaccine, in particular to a method for preparing an inactivated rabies vaccine for animals using a bioreactor. Background technique [0002] Rabies, commonly known as "hydrophobia", is a zoonotic infectious disease of the central nervous system caused by rabies virus. Its clinical manifestations are mainly characterized by hydrophobia, photophobia, dysphagia, and mania. 100% acute infectious disease. The "Law of the People's Republic of China on the Prevention and Control of Infectious Diseases" lists rabies as a Class B infectious disease. [0003] In recent years, the number of dogs and cats in my country has increased rapidly, and the number of people injured by dogs and cats has also continued to increase. According to the amount of human rabies vaccine used in my country, it is estimated that more than 40 million people are injured by animals every year in the country. Rabies deaths accounted f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/135C12N7/00
Inventor 岳雷李淑芬陈静郭鑫韩中山张振山
Owner TANGSHAN YIAN BIOLOGICAL ENG CO LTD
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