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Method for preparing vaccine by culturing varicella virus with microcarriers

A microcarrier culture, varicella virus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, viruses, etc., can solve the problem of excessive bovine serum residual volume, achieve high start-up time and production efficiency, control residual volume, Produce fast effects

Active Publication Date: 2018-04-13
CHANGCHUN KEYGEN BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Solved the technical problem that the residual amount of bovine serum in the microcarrier culture method of the bioreactor exceeded the standard

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Pretreatment of microcarriers. 0.05M pH9.6 carbonate buffer was used to prepare recombinant human serum albumin with a concentration of 2% as a coating solution; polystyrene microcarriers were soaked in the coating solution and coated at 4°C for 8h.

[0031] Seed cell culture. Cultivate human diploid cells 2BS on the wall with MEM medium containing 10% newborn bovine serum through cell culture flasks. When the confluence of the cell monolayer reaches 80-90%, the cells that pass the microscope inspection are treated with 0.25% trypsin Digest to prepare cell suspension as seed cells.

[0032] Culture of production cells. In the bioreactor, add freshly prepared aseptic MEM cell culture fluid, pretreated microcarriers, and 10% neonatal bovine serum in total volume, wherein the concentration of polystyrene microcarriers is 3-6g / L. The seed cell suspension was inoculated into the bioreactor at an inoculation concentration of 1×10 4 -1×10 5 cell / mL, under the conditions o...

Embodiment 2

[0037] Except in the microcarrier washing step, the microcarriers were washed 3 times with 0.01M pH7.4 PBS. After testing, the concentration of bovine serum albumin in the buffer solution after washing was about 30ng / mL. The specific operations and conditions of the other steps Parameters are all the same as in Example 1.

Embodiment 3

[0039] Except in the varicella virus inoculation step, the working passage of the varicella-zoster virus vaccine strain with a virus titer of about 5.0 lgPFU / mL was used to inoculate the varicella-zoster virus at a multiplicity of infection (MOI) of 0.008 relative to cells Except that, the specific operations and condition parameters of the remaining steps are all the same as those in Example 1.

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Abstract

The invention relates to a method for preparing a vaccine through microcarrier culture of varicella-zoster viruses. The method comprises the following steps: implementing pre-treatment on a microcarrier; implementing culture of seed cells; implementing culture of production cells; washing the microcarrier; implementing varicella-zoster virus vaccination; and preparing the vaccine. The method provided by the invention, by implementing the coating pre-treatment on the microcarrier and by washing the microcarrier before the virus vaccination, can greatly reduce serum residues and improve titer of a virus stock solution.

Description

technical field [0001] The present invention relates to the production technology of biological product, be specifically related to a kind of production preparation method of varicella virus vaccine, relate in particular to a kind of method that utilizes bioreactor microcarrier to cultivate cells, produce varicella virus stock solution to prepare attenuated live vaccine and its product. Background technique [0002] Chickenpox is an acute respiratory infectious disease caused by varicella-zoster virus (VZV). It is highly contagious and has obvious infectivity and pathogenicity to children. Vesicle rash. Simple varicella virus infection can heal itself, but if it is not handled properly, it is easy to cause severe diseases such as pneumonia and encephalitis, and it can also produce herpes zoster if it is latent in the body after infection. Inoculation of attenuated live varicella vaccine is an effective means to control the infection and epidemic of varicella. At present, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/25A61P31/22C12N7/00C12R1/93
CPCA61K39/12C12N7/00C12N2710/16734C12N2710/16751
Inventor 李春明徐俊峰朱晓文王磊闫磊赵海波双慧王玮沈红杰
Owner CHANGCHUN KEYGEN BIOLOGICAL PROD
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