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Bioreactor process of chick embryo cell rabies virus

A bioreactor, rabies virus technology, applied in embryonic cells, viruses, microorganisms, etc., can solve the problems of low culture cell density, high labor intensity, low virus titer, etc., and achieve high culture density, high safety, and operation. simple effect

Active Publication Date: 2021-02-09
SHENZHEN WEIGUANG BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent of Chinese Patent Publication No. CN108452298A discloses a process for producing live attenuated yellow fever vaccine with SPF chicken embryo cells, which has the disadvantages of low culture cell density, low virus titer, and high labor intensity.
The Chinese patent publication number is CN1390604, which discloses a method for large-scale continuous production of virus vaccines. The CelligenPlus bioreactor with a fixed bed basket stirring system is used, and the polyester chip Fibra-Cel Disks is used as a carrier, and the scale is only 5.0 -7.5L, not suitable for large-scale rabies vaccine production

Method used

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  • Bioreactor process of chick embryo cell rabies virus
  • Bioreactor process of chick embryo cell rabies virus
  • Bioreactor process of chick embryo cell rabies virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] (1) Choose 9-11 days old chicken embryos with normal development, visible blood vessels and activities. Soak in 0.2% (m / v) bromogeramine solution for 5-10 minutes, then spray and cauterize with 75% (v / v) alcohol. Aseptically take out the chicken embryos and put them into a plate filled with PBS solution (pH7.4). Remove the head of the chicken embryo, put it into a sterilized jar, and cut it into 1~5mm with sterile scissors 3 Add 0.25% (m / v) trypsin solution preheated to 37°C according to the amount of 5-8mL per chicken embryo, and place it in a 37°C water bath for 15-30 minutes for digestion. Centrifuge at room temperature at 1500-3000 rpm for 8-10 minutes, remove the supernatant, and resuspend the precipitate with serum-free medium to prepare a single chicken embryo cell suspension. Take 2mL of cell suspension and add 16mL of purified water to mix evenly, read with a turbidimeter, and calculate the number of cells according to the following formula:

[0054] Cell de...

Embodiment 2

[0060] The difference with Example 1 is that the cell seeding density when the bioreactor cultivates cells is 3.0×10 6 individual / mL.

Embodiment 3

[0062] The difference with Example 1 is that the cell seeding density when the bioreactor cultivates cells is 3.6×10 6 individual / mL.

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Abstract

The invention relates to a bioreactor process of a chick embryo cell rabies virus. Primary chick embryo cells are cultured in a bioreactor with a fixed bed basket type stirring system by using a sheetcarrier and a serum-free culture medium, and rabies virus harvesting liquid with high titer and high antigen is prepared through specific in-tank cell density, inoculation amount of CTN chick embryocell vaccine strains-CTNCEC25 strains (CGMCC No.6510) and set parameters of the bioreactor. The virus titer is averagely not lower than 7.0 lgFFU / mL, the highest virus titer can reach 8.7 lgFFU / mL, the antigen content is averagely not lower than 2.0 IU / mL, the highest antigen content can reach 4.0 IU / mL, the harvesting liquid can reach 2-3 effective culture volumes, the labor intensity and the production cost are greatly reduced, the production efficiency is greatly improved, and a culture mode for producing rabies viruses through primary cell large-scale culture is created.

Description

technical field [0001] The invention relates to the technical field of biological products and is applied to the production of primary chicken embryo cell rabies vaccine, in particular to a bioreactor process for chicken embryo cell rabies virus. Background technique [0002] Rabies (Rabies) is an ancient and severe human-animal infectious disease caused by Rabies virus (RABV) that attacks the central nervous system and can cause severe encephalomyelitis. Once the disease occurs, the mortality rate is almost 100%. Almost all warm-blooded animals can be infected with rabies, especially with various mammals as their main hosts. There is currently no effective treatment for rabies, and timely rabies vaccination after exposure is the most important means of prevention. [0003] After more than 100 years of development, human rabies vaccines have developed from the earliest attenuated active vaccines to poultry embryo vaccines to the current cell culture vaccines and genetic eng...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N5/02C12N7/00C12N7/02
CPCC12N5/0603C12N7/00C12N2531/00C12N2509/00C12N2509/10C12N2500/90C12N2501/998C12N2501/90C12N2760/20151
Inventor 罗姗李慧王春华周维陈楚如吴灼斌洪玮琪黄致翔曹静梁琳琳郭采平黄伟荣李云富刘永娣刘璐谭梦思刘爱和李彬梁富徐尔赞刘依润
Owner SHENZHEN WEIGUANG BIOLOGICAL PROD
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