Method for producing influenza vaccine in large-scale by using bioreactor

A technology of bioreactor and influenza vaccine, which is applied in the field of large-scale production of influenza vaccine with bioreactor, can solve the problems of influenza virus production of influenza vaccine, etc., and achieve the effects of easy scale expansion, high antigenic content, and large reaction

Inactive Publication Date: 2007-10-31
江苏全益生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because in the cell culture technique, the virus is adsorbed to the cells to reproduce, the above-mentioned rabies, hemorrhagic fever, Japanese encephalitis, and polio viruses are easily adsorbed on the cells, so the above method is suitable for rabies vaccines, hemorrhagic fever vaccines, The preparation of Japanese encephalitis vaccine, polio vaccine and rotavirus vaccine, and influenza virus is difficult to produce influenza vaccine by the method proposed in the above-mentioned patent without Vero cell adaptation, and this is that so far influenza vaccine still relies on chicken embryo Reasons why large-scale production of influenza vaccines cannot be produced in bioreactors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] The influenza vaccine is prepared as follows:

[0013] In the preparation of influenza vaccine, the first thing to be solved is that influenza virus adapts to Vero cells, which is the key to this preparation method. The following steps are taken: recover a Vero cell from the seed cell bank, and add the cells to cells with growth fluid In the culture vessel, the growth medium is MEM or M199+8% bovine serum pH 7.0-7.2. After culturing at 37°C for 48 hours to grow into a single layer, discard the growth medium, wash the cell surface 1-2 times with Hank's solution, and place Wash the remaining bovine blood in the growth solution (because bovine serum contains non-specific inhibitors of influenza virus, which will affect the replication of influenza virus), then inoculate influenza virus seeds, place at 33-35°C for 1-2 hours, and then add Maintenance solution, the maintenance solution adopts serum-free medium (containing 1-5 μg / ml trypsin), and the purpose of adding trypsin ...

Embodiment 2

[0019] The bioreactor adopts a suspension culture reactor, the carrier is a dextran polymer, and the operation steps are basically the same as in Example 1.

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PUM

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Abstract

The invention discloses a method to mass produce flu vaccine with bioreactor, which comprises the following steps: proceeding passage adaptation on Africa green monkey kidney cell (Vero cell) with influenza virus; intensifying intrinsic appeal of influenza virus for Vero cell; fitting for Vero cell quickly; breeding the Vero cell with cell density above 10. 7deka / ml in bioreactor; seeding the influenza virus into bioreactor; adjusting various controlling parameter in the reactor; making the virus high effective breed in the reactor; proceeding continuous perfusion; harvesting virus liquid; deactivating the virus; condensing; purifying; cracking; getting the flu vaccine. This invention possesses the advantages of high antigen quantity and stable quality.

Description

technical field [0001] The invention relates to a method for large-scale production of influenza vaccine using a bioreactor. Background technique [0002] The influenza vaccine that has been approved for production is still an inactivated vaccine derived from chicken embryos, but it still has many deficiencies, such as, the continuous passage of chicken embryos often causes strain variation; chicken embryos may be contaminated by exogenous virus factors; Chicken embryo production of influenza vaccine can not meet the needs of emergency situations, especially the outbreak of bird flu, will also affect the normal supply of chicken embryos. Therefore, the World Health Organization is urging countries around the world to find ways to mass-produce influenza vaccines from cell lines sensitive to influenza viruses. Chinese Patent No. 02112013.7 discloses a method for large-scale continuous production of virus vaccines in the specification for a patent for invention, which comprise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/145A61P31/16C12M3/02C12M1/02
Inventor 张玉慧
Owner 江苏全益生物科技股份有限公司
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