Diploid cell cerebritis B vaccine and purified cerebritis B vaccine, dosage form freeze-drying and water injection

A diploid cell and Japanese encephalitis technology, which is applied in the direction of freeze-drying delivery, antiviral agent, drug delivery, etc., to achieve the effect of improving purity, improving stability and increasing persistence

Active Publication Date: 2005-03-02
崔栋
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But prepare Japanese encephalitis vaccine with this method and there is no report

Method used

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  • Diploid cell cerebritis B vaccine and purified cerebritis B vaccine, dosage form freeze-drying and water injection

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Experimental program
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Effect test

Embodiment 1

[0053] Diploid cells come from the Chinese Center for Disease Control and Prevention, use 38-50 generations, the cell nutrient solution is 199 medium containing 2-10% bovine serum and 25IU / ml gentamicin and 25IU / ml kanamycin, and Adjust the pH to 7.2, cultivate a dense monolayer with a 3L spinner bottle at 37°C, and then amplify according to the seeding ratio of 1:2. When the expansion reaches the production batch, after about 4 days, when the cells grow into a dense monolayer, discard the cells Nutrient solution, wash the cell surface with PH7.2 Earl's solution, inoculate diploid cells with JE virus, and use JE mouse encephalovirus P 3 The second-generation working virus strain (P 3-2 ), virus seed concentration MOI0.05, cell maintenance medium is 199 culture medium containing 0.4-0.5% (W / W) human albumin, pH 7.6, cultivated at 35°C, after 24 hours, replace with fresh cell maintenance medium to continue Cultivate for 2 days, start to harvest the virus, harvest once every 3-4...

Embodiment 2

[0057] Diploid cells come from the Chinese Center for Disease Control and Prevention, use 38-50 generations, the cell nutrient solution is 199 medium containing 2-10% bovine serum and 25IU / ml gentamicin and 25IU / ml kanamycin, and Adjust the pH to 7.2, cultivate a dense monolayer with a 3L spinner bottle at 37°C, and then amplify according to the seeding ratio of 1:2. When the expansion reaches the production batch, after about 4 days, when the cells grow into a dense monolayer, discard the cells Nutrient solution, wash the cell surface with PH7.2 Earl's solution, inoculate diploid cells with Japanese encephalitis virus, and use the second generation of JE virus strain AS14-14-2 to pass on diploid cells Poison species (P 3-2 ), virus seed concentration MOI0.05, cell maintenance medium is 199 culture medium containing 0.4-0.5% (W / W) human albumin, pH 7.6, cultivated at 35°C, after 24 hours, replace with fresh cell maintenance medium to continue Cultivate for 2 days, start to ha...

Embodiment 3

[0061] The JE inactivated vaccine prepared by the method of claim 1 is tested (including giving injections to 12 children aged 4-6 years, and those who have not been vaccinated against JE vaccine are inoculated with 2 needles of JE inactivated vaccine, with an interval of 7-10 days between the two needles. .) proved that the positive conversion rate of neutralizing antibodies was 91.7%, and no side effects occurred.

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Abstract

The present invention relates to cerebritis B vaccine of diploid cell and purified cerebritis B vaccine in the preparation form of freeze dried powder for injection and injection liquid. The purified cerebritis B vaccine is obtained through inoculating cerebritis B virus strain onto diploid cell for human body.

Description

Technical field: [0001] The invention relates to a purified Japanese encephalitis vaccine, in particular to a purified Japanese encephalitis vaccine obtained by inoculating Japanese encephalitis virus seeds on human diploid cells. Background technique: [0002] At present, there are three kinds of Japanese encephalitis vaccines produced in large quantities in the world. The first one is a purified Japanese encephalitis vaccine prepared by grinding, emulsifying, concentrating, and ultracentrifuging the brain tissue of mice infected with JE virus, represented by Japan; It is the inactivated JE vaccine cultured from primary hamster kidney cells and the live JE vaccine from primary hamster kidney produced in China. The cell substrates of these two vaccines are derived from ordinary animals, and there is no guarantee that they will not carry foreign factors; The third is the freeze-dried JE purified and inactivated vaccine produced by the Beijing Institute of Biological Products ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/08A61K9/19A61K39/12A61P31/12
Inventor 崔栋卜海兵
Owner 崔栋
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