Diploid cell rabies vaccine and method for preparing purified rabies vaccine

A technology of diploid cells and vaccines is applied in the field of preparation of purified rabies vaccines, which can solve the problems that exogenous factors cannot be guaranteed, that cells cannot be carried, and the tumorigenicity of cell matrix and cell DNA cannot be guaranteed, so as to achieve good antibodies. Level, stability improvement, effect of increasing persistence

Inactive Publication Date: 2009-01-28
崔栋
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, there are four kinds of rabies vaccines mass-produced in the world: the first kind is diploid cells represented by the United States, which is prepared by concentration and ultracentrifugation; Dry inactivated vaccine, but does not guarantee tumorigenicity of the cell matrix and cellular DNA
The third and fourth types of vaccines are hamster kidneys represented by China and chicken embryos and duck embryos from Japan. The cells of these types of vaccines are derived from common animals. It cannot be guaranteed that the cells do not carry foreign factors and the tumorigenicity and tumorigenicity of the cell matrix cannot be guaranteed. cell DNA

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Diploid cells come from the Chinese Center for Disease Control and Prevention, and the serum-free medium of the cell nutrient solution neutralizes 25IU / ml gentamicin or 25IU / ml kanamycin, and adjusts the pH to 7.2. After the dense monolayer was amplified according to the seeding ratio of 1:2, when it was amplified to the production batch, after about 4 days, when the cells grew into a dense monolayer, the cell nutrient solution was discarded, and the SNK-CTN virus was inoculated with a virus seed concentration of MOI 0.1~0.01, after 3 days of cultivation, start to harvest the virus, harvest once every 3-4 days, and collect 3 times in total, each batch of virus liquid is sampled for virus titration and sterility test, and it is required to harvest the virus drops of the virus liquid for the virus liquid degrees up to 10 5.0 LD50 / ml or above; add formaldehyde solution or β-propiolactone (final concentration 1 / 4000) or other suitable inactivating agents to the combined vir...

Embodiment 2

[0059] Diploid cells come from the Chinese Center for Disease Control and Prevention, and the cell nutrient solution is serum-free medium neutralized with 25IU / ml gentamicin or 25IU / ml kanamycin, and adjust the pH to 7.2, culture in a 3L spinner bottle at 37°C After forming a dense monolayer, amplify according to the seeding ratio of 1:2. When the amplified to the production batch, after about 4 days, when the cells grow into a dense monolayer, discard the cell nutrient solution, inoculate diploid cell rabies virus, and use Rabies virus SNK-CTN. The second-generation working virus seed of the strain passed on diploid cells, the concentration of the virus seed is MOI 0.05, and the cell maintenance medium is 199 culture medium containing 0.4-0.5% (W / W) human albumin, pH7.6, 35°C After 24 hours, replace with fresh cell maintenance medium and continue to cultivate for 2 days. Harvest the virus every 3.4 days, a total of 3 times. Each batch of virus liquid is sampled for virus titr...

Embodiment 3

[0063] With the inactivated rabies vaccine prepared by the method of claim 1, after 14 days of inoculating the purified rabies vaccine to those who have not been vaccinated against rabies, the positive conversion rate of neutralizing antibodies is higher than 95.0%, and no side effects occur.

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Abstract

The invention relates to a preparation method for a diploid cell rabies vaccine and a purified rabies vaccine, and the method uses a serum-free medium to culture human diploid cell rabies purified vaccine, thus greatly reducing anaphylactic reaction and being beneficial to purification.

Description

Technical field: [0001] The invention relates to a preparation method of a purified rabies vaccine, in particular to a preparation method of a purified rabies vaccine obtained by inoculating rabies virus seeds on human diploid cells. Background technique: [0002] At present, there are four kinds of rabies vaccines mass-produced in the world: the first kind is diploid cells represented by the United States, which is prepared by concentration and ultracentrifugation; Dry inactivated vaccines, but can not guarantee the tumorigenicity of the cell matrix and the DNA of the cells. The third and fourth types of vaccines are hamster kidneys represented by China and chicken embryos and duck embryos from Japan. The cells of these types of vaccines are derived from ordinary animals. It cannot be guaranteed that the cells do not carry exogenous factors, nor can the tumorigenicity and tumorigenicity of the cell matrix. the DNA of the cell. Chinese patent 200510080058.2 describes a pur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/205A61P31/14C12N5/08C12N7/02
Inventor 崔栋
Owner 崔栋
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