Method for producing rabies virus antigens for animals at a large scale

A rabies virus, large-scale technology, applied in the field of veterinary biological products, can solve the problems of no special and effective treatment for rabies, large differences between batches, unstable quality, etc., to save costs and labor, easy control, and reduce production. cost effect

Inactive Publication Date: 2010-10-06
PU LIKE BIO ENG
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Problems solved by technology

However, there is no specific and effective treatment for rabies, and rabies vaccination is the only effective method
[0003] At present, the rabies virus vaccines on the market are basically subcultured by the spinner bottle process. Large-scale production requires a lot of material and human resources, and the cost is high, and the difference between batches is large, and the quality is unstable.

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  • Method for producing rabies virus antigens for animals at a large scale

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Embodiment 1

[0032] 1. Viruses and cell lines

[0033] The virus strain used to prepare the rabies virus antigen is the LEP-Flury strain, which has no pathogenicity after pathogenicity test. The hamster kidney cell (BHK21), a cell line with good sensitivity to the virus strain, is used as the cell line for vaccine production, so as to infect and multiply the rabies virus in large quantities.

[0034] 2. Preparation method

[0035] (1) Inoculate the cell line hamster kidney cells (BHK21) for the preparation of antigen into DMEM liquid medium (including 5% bovine serum, 0.01mol / L NaHCO 3 , 0.1mg / ml kanamycin sulfate (kanamycinsulfate), 100,000IU penicillin G sodium salt (Penicillin G Sodium), pH value is 7.2) and in the carrier tank of BioNoc II polyester fiber; Add 5.5 g of microcarriers, and the initial inoculum size of cells is 2.7 × 10 7 cells / g microcarriers.

[0036] (2) At 37°C, 5% CO 2 Under the culture environment, the cells are attached for 4 hours, the above-mentioned cells f...

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Abstract

The invention discloses a method for producing rabies virus antigens for animals at a large scale, which produces the rabies virus antigens at a large scale by utilizing a bioreactor by a cell microcarrier suspension culture system. The method comprises the following steps: inoculating cells for preparing the antigens into a carrier tank containing a culture solution and microcarriers to enable the cells to be attached to the microcarriers; in a proper culture environment, enabling the cells to grow on the microcarriers until the quantity of the cells is 5-40 times more than inoculum density; making rabies viruses into a virus suspension, and enabling the virus suspension to be adsorbed on the cells; culturing the viruses in the proper culture environment by using a cell maintenance culture solution; continuously culturing for 3-5 days and then harvesting a virus solution for the first time, wherein a semicontinuous process is adopted and the ratio of a changed solution is 50 percent; continue culturing for 9-11 days, and harvesting the changed solution once every 24 hours; mixing the harvested virus solution with the virus solution of the bioreactor; and carrying out freeze thawing at the temperature of -20 DEG C and inactivation purification to obtain the rabies virus antigens. The method has large production scale, high single-scale yield and relatively low production cost.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, relates to animal viruses, and more specifically relates to a method for producing rabies virus antigens for animals. Background technique [0002] Rabies is a zoonotic disease caused by rabies virus (Rabies virus, RV) infection with severe symptoms and high fatality rate. The mortality rate is almost 100%. Currently, there is no effective treatment. Wild animals are the main storage hosts of RV. Once people are bitten by such animals carrying rabies virus, they will be infected and diseased, and the case fatality rate is 100%. According to the latest statistics on infectious disease epidemics released by the Ministry of Health, it shows that rabies continues to show signs of danger in recent years. However, there is no specific and effective treatment for rabies, and rabies vaccination is the only effective method. [0003] Rabies virus vaccines currently on the market a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/02
Inventor 张许科孙进忠乔荣岑陶家权王进产张海洋习向锋李三
Owner PU LIKE BIO ENG
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