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Vaccine for virus of encephalitis B and preparation method

A Japanese encephalitis virus and vaccine technology, applied in antiviral agents, virus antigen components, pharmaceutical formulations, etc., can solve the problems of inability to completely remove animal-derived exogenous factor contamination, tumorigenic phenomena, etc., and achieve good protection effect , High immunogenicity and potency, Acceptable effect

Active Publication Date: 2005-11-16
复星安特金(成都)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of these vaccines are: firstly, the contamination of exogenous factors of animal origin cannot be completely removed
Although the safety of the inactivated vaccine prepared by Vero cell culture has been significantly improved, there is tumorigenic phenomenon in nude mice after Vero cells exceed 232 passages (Levenbook IS, et al, Tumorigenicity of vero cells.J Biol Stand 1984 Oct ;12(4):391-8)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of human diploid cells

[0034] The preparation of human diploid cells takes MRC-5 as an example: the cell culture medium is DMEM (other suitable cell nutrient solutions such as MEM, 199 medium, 1640 medium, etc. can also be used), and 4-10% fetal bovine serum is added (No exogenous factor contamination) and kanamycin (or gentamicin) with a concentration of 100-200U / ml, adjust the pH to between 7.0-8.0. Cells are cultured at 35-37°C with Koshik flasks or roller bottles. After 4-8 days, they are cultured into dense monolayer cells, and then expanded at a ratio of 1:2 until the production batch is expanded. After 3-5 days After culturing, the cells grow into dense monolayer cells, and the cell culture medium is discarded. The surface of the cells was rinsed with phosphate buffer solution of pH 7.0-8.0 to wash away the residual bovine serum. Prepare for virus inoculation.

Embodiment 2

[0036] Domestication, adaptation and expansion of Japanese encephalitis virus in human diploid cells

[0037] Take human diploid cells and MRC-5 as an example: because the virus reproduction titer of JE virus in human diploid cells is low, it is necessary to gradually domesticate and adapt JE virus in human diploid cells. Inoculate the Japanese encephalitis virus P3 strain (or other virus strains such as SA14) into the human diploid cells that have grown to a monolayer, and continue to culture at 30-37°C. When the cytopathy reaches 50-70%, the cell supernatant is harvested, which is the Japanese encephalitis virus liquid that grows and reproduces in the human diploid cells. Successive generation-to-generation adaptations are carried out. When the Japanese encephalitis virus titer in the harvested Japanese encephalitis virus liquid reaches more than 7.5lgPFU / ml, it is used as the seed virus for the production of Japanese encephalitis vaccine. According to the requirements of ...

Embodiment 3

[0039] Passage Stability of Japanese Encephalitis Virus in Human Diploid Cells

[0040] Carry out successively by the test procedure of embodiment 2, can observe that the virus titer of Japanese encephalitis virus in human diploid cell improves gradually, more than or equal to 7.5lgPFU / ml, and tends towards stability, the virus titer of each generation is shown in the table 1.

[0041] Virus Adaptation Generation

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PUM

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Abstract

A vaccine of the encephalitis B virus for preventing viral encephalitis B is prepared through inoculating said virus into human diploid cells, culturing, naturalizing, adapting, amplifying, concentrating and purifying.

Description

technical field [0001] The invention relates to a Japanese encephalitis virus vaccine prepared by human diploid cell culture. Specifically, the present invention relates to a method of inoculating Japanese encephalitis virus in human diploid cells for cultivation, and after domestication, adaptation and amplification processes, the Japanese encephalitis virus reproduces in large numbers in human diploid cells, It is then concentrated and purified to prepare a Japanese encephalitis virus vaccine. The virus vaccine can prevent Japanese encephalitis disease caused by Japanese encephalitis virus infection in infants, children, teenagers and adults. The present invention also relates to the preparation method of this virus vaccine. Background technique [0002] Japanese encephalitis is an infectious disease caused by acute viral infection of the central nervous system. The annual incidence rate in endemic areas is as high as 100-100 / 100,000. Nearly 3 billion people live in Japa...

Claims

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Application Information

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IPC IPC(8): A61K39/12
Inventor 薛平
Owner 复星安特金(成都)生物制药有限公司
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