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Method for mass production for recombinant adenovirus for novel coronavirus gene vaccine

A technology of recombinant adenovirus and coronavirus, applied in biochemical equipment and methods, viruses, genetic engineering and other directions, can solve problems such as the inability to meet the requirements of large-scale industrialization of gene therapy products, and achieve the effect of high preparation efficiency

Pending Publication Date: 2020-07-10
SYNO SHENZHEN BIOMEDICAL RES CO LTD
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AI Technical Summary

Problems solved by technology

Traditional laboratory gene therapy product preparation methods can no longer meet the requirements of GMP large-scale industrialization of gene therapy products

Method used

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  • Method for mass production for recombinant adenovirus for novel coronavirus gene vaccine
  • Method for mass production for recombinant adenovirus for novel coronavirus gene vaccine
  • Method for mass production for recombinant adenovirus for novel coronavirus gene vaccine

Examples

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Effect test

Embodiment 1

[0036] Embodiment 1: Production process development of recombinant adenovirus vector gene vaccine product

[0037] In this example, the production of a recombinant adenovirus vector gene vaccine for the prevention of novel coronavirus is illustrated as an example. The production process specifically includes the following steps, all of which are carried out in a GMP clean room.

[0038] 1. Cell Expansion

[0039] Take 1 cell of the recombinant adenovirus working cell bank, which contains the recombinant adenovirus vector gene used to prevent the new coronavirus, and the cell volume is about 5x10 6 to 1x10 7 , cells were thawed and inoculated into T-flasks for cell culture at a seeding density of about 0.5x10 4 to 5x10 4 pieces / cm 2 . Cultivate in a 5% carbon dioxide incubator at a temperature of 37°C until the cell density reaches about 90%, and harvest the cells by detaching them with trypsin-EDTA.

[0040] Inoculate the harvested cells into more T-flasks to expand the ...

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Abstract

The invention discloses a method for mass production for a recombinant adenovirus for a novel coronavirus gene vaccine. The method comprises the following steps of carrying out cell proliferation; inoculating cells harvested through proliferation into a packed bed bioreactor for culture; carrying out virus infection on the cells in the packed bed bioreactor and harvesting a culture solution containing a recombinant adenovirus; filtering and concentrating a recombinant adenovirus collection fluid; adding broad-spectrum nuclease into the recombinant adenovirus collection fluid and carrying out nuclease treatment; and separating and purifying the recombinant adenovirus collection fluid subjected to nuclease treatment by using an anion exchange column chromatography and obtaining the purifiedrecombinant adenovirus. Infection and proliferation of the recombinant adenovirus are carried out by adopting the packed bed bioreactor, so that the efficiency is high and the method is more suitablefor the requirements of a GMP production environment. Separation and purification are carried out by adopting full-automatic column chromatography, so that more economical and efficient large-scale industrial GMP production of recombinant adenovirus preparation products is achieved, and the novel coronavirus gene vaccine is used for preventing infection and transmission of a novel coronavirus.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceutical technology, and relates to a method for mass production of recombinant adenoviruses for novel coronavirus gene vaccines, which is applied to the mass production process of recombinant adenovirus gene therapy products GMP, especially for novel coronaviruses GMP mass production process of recombinant adenovirus gene vaccine. Background technique [0002] After decades of research and development, the field of gene therapy has made great progress and achieved product breakthroughs. The recombinant adenovirus p53 gene therapy product of Shenzhen Saibainuo Gene Technology Co., Ltd., the Glybera recombinant adeno-associated virus (AAV) product of the Netherlands UniQure company and the Luxturna recombinant adeno-associated virus (AAV) product of the US Spark Therapeutics company have been approved by the government successively. Approved by the pharmaceutical regulatory agency for marketing. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861
CPCC12N15/86C12N2710/10343C12N2710/10352
Inventor 徐卫
Owner SYNO SHENZHEN BIOMEDICAL RES CO LTD
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