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Method for culturing CHO cells on large scale to prepare hepatitis B vaccine

A large-scale culture and hepatitis B technology, applied in the direction of biochemical equipment and methods, animal cells, tissue culture, etc., can solve the problems of high cost, low efficiency, easy pollution, etc., to improve production efficiency and overcome low production efficiency , The effect of overcoming the risk of easy pollution

Inactive Publication Date: 2020-01-17
苏州百奥思达生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, mammalian cells have very high requirements on culture conditions. The existing production process is a culture process using spinner bottles, which has the risks of low efficiency, high cost, and easy pollution.

Method used

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  • Method for culturing CHO cells on large scale to prepare hepatitis B vaccine
  • Method for culturing CHO cells on large scale to prepare hepatitis B vaccine

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Embodiment Construction

[0027] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0028] refer to figure 1 with figure 2 , a method for large-scale culture of CHO cells in the first embodiment of the present invention to prepare hepatitis B vaccine, comprising the following steps,

[0029] S1. Cell recombination: provide recombinant CHO cells with HBsAg antigen sequence, and carry out passage and increase of the recombinant CHO cells under the condition that the culture environment temperature is 35° C., and the culture time is 6 days.

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Abstract

The invention discloses a method for culturing CHO cells on large scale to prepare a hepatitis B vaccine. The method comprises cell recombination, cell proliferation, perfusion culture, concentrated enrichment of a collected culture solution, purification, addition of formaldehyde into a purification solution for treatment, preparation of a semi-finished product and packaging of the semi-finishedproduct to obtain a finished product. The perfusion culture comprises specific steps as follows: when the cell density is proliferated to 2*10<6> cells / ml or above, a maintenance medium is replaced for perfusion culture, during perfusion culture, the cell density after culture can reach 3*10<7> cells / ml, the motility rate is larger than 98%, the HBsAg antigen field corresponding to a collected solution can reach 13 mg / L or above, 50% higher than an existing spinner bottle process, so that the production efficiency of the recombinant hepatitis B vaccine, and the defect of low production efficiency in the prior art is overcome.

Description

technical field [0001] The invention relates to the field of vaccine production, in particular to a method for large-scale culture of CHO cells to prepare hepatitis B vaccine. Background technique [0002] Existing hepatitis B vaccines include yeast hepatitis B vaccine and CHO cell hepatitis B vaccine. Both vaccines are genetically recombined biological products made using modern biotechnology. [0003] Yeast recombinant hepatitis B vaccine is a vaccine containing HBsAg subunits. It uses modern biological technology to clone the HBsAg gene into yeast, and expresses hepatitis B surface antigen by cultivating recombinant yeast. Although the yeast system has the advantages of high expression and low cost, it has the disadvantage of aglycosylation of HBsAg produced by the expression system, and it cannot be secreted, and the separation and purification of the product is difficult. [0004] CHO cell hepatitis B vaccine is to clone HBsAg gene into CHO cells, and express hepatiti...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/29A61P31/20C12N5/10C12P21/02
CPCA61K39/12A61P31/20C07K14/005C12N5/0682C12N2510/02C12N2730/10122C12N2730/10134C12N2730/10151
Inventor 赵永强
Owner 苏州百奥思达生物科技有限公司
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