Human diploid cell rabies vaccine and preparation method thereof

A technology of human diploid cells and rabies vaccine, which is applied in the field of bioengineering, can solve the problems of expensive vaccines, low virus titers, HDCs are not easy to cultivate, etc., and achieve vaccines with less impurities, perfect purification process, and high purity. The effect of high immunogenicity

Active Publication Date: 2012-09-19
CHENGDU KANGHUA BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are four kinds of rabies vaccines mass-produced in the world: the first kind is the human diploid cell represented by the United States, and the rabies vaccine prepared by concentration and ultracentrifugation; the second kind is Vero cell represented by France as the culture substrate, produced Freeze-dried inactivated vaccines, but the tumorigenicity of the cell matrix and the safety of cellular DNA cannot be guaranteed
The third and fourth types are hamster kidneys represented by China and chicken embryos and duck embryos from Japan; these types of vacci

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Firstly, human diploid cell WI-38 is recovered, cultured and expanded to meet the needs of production batches. Animal cell culture medium 199 culture medium was used, 2% bovine serum and kanamycin 26U / ml were added to the cell culture medium, pH 7.0, the culture temperature was 37°C, and the culture method was roller bottle culture.

[0053] During the culture process, the cell separation rate is determined according to the needs of the production batch. In this example, 1:2 is selected. When the cells grow into a dense monolayer, the rabies virus can be inoculated, and 0.3% (W / W) is added to the cell maintenance solution. At the same time, adjust the pH to 7.0, culture at 37°C, and add an appropriate amount of antibiotics. When inoculating, first wash the cell surface with a culture medium such as Earle's solution to remove residual bovine serum, and then grow dense cells Inoculate 0.1 mol of rabies virus strain CDKHBP-1 strain on the human diploid cells of the layer, ...

Embodiment 2

[0058] Firstly, human diploid cell WI-38 is recovered, cultured and expanded to meet the needs of production batches. Animal cell culture medium 199 culture medium was used, 10% bovine serum and gentamicin 30U / ml were added to the cell culture medium, pH 7.5, the culture temperature was 35°C, and the culture method was microcarrier reactor culture.

[0059] During the culture process, the cell sorting rate is determined according to the needs of the production batch. In this embodiment, 1:4 is selected. When the cells grow into a dense monolayer, the rabies virus can be inoculated. It is best to add 0.5% (W / W) human serum albumin, adjust the pH to 7.5 at the same time, culture at 35°C, and add an appropriate amount of antibiotics. The culture medium that can be used includes 199 liquid, constant salt solution, etc. The inoculation process includes first using a culture medium such as Earle's solution Wash the cell surface to remove residual bovine serum, then inoculate 0.01 m...

experiment example

[0065] After the rabies vaccine prepared by the method of the present invention is given to people who have not been vaccinated against rabies vaccine for 15 days, the positive conversion rate of neutralizing antibodies is higher than 98%, and no side effects occur.

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Abstract

The invention discloses a human diploid cell rabies vaccine and a preparation method thereof, and belongs to the field of vaccines. A CDKHBP-1 strain is inoculated into a human diploid cell WI-38, and the rabies vaccine is obtained through separation and purification. The human diploid cell does not contain any contaminants or oncogenicity, and is obvious superior to an animal cell serving as a medium for producing the vaccine; and the method is suitable for large-scale industrial production, a purification process is perfect, and the vaccine contains a few impurities and has high purity and immunogenicity.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a human diploid cell inactivated vaccine, in particular to a rabies inactivated purified vaccine obtained by inoculating human diploid cells with a rabies virus strain and a preparation method thereof. Background technique [0002] Rabies, rabies, also known as hydrophobia, is an acute viral infectious disease that affects the central nervous system. All warm-blooded animals, including humans, may be infected. It is mostly obtained from the bite of infected animals. Since there is no effective treatment for rabies and the mortality rate is extremely high, vaccination is the only effective means to prevent the occurrence of rabies and ensure people's health. [0003] At present, there are four kinds of rabies vaccines mass-produced in the world: the first kind is the human diploid cell represented by the United States, and the rabies vaccine prepared by concentration and ultracentri...

Claims

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Application Information

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IPC IPC(8): A61K39/205A61P31/14C12N7/00C12N7/02C12R1/93
Inventor 蔡勇侯文礼钟泽荣冯晓
Owner CHENGDU KANGHUA BIOLOGICAL PROD
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