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Method for producing rabies vaccine by applying bioreactor and sheet carrier

A bioreactor, rabies vaccine technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of long production cycle, low virus harvest, unsuitable for large-scale production, etc., and achieve production costs Effect of low, high cell density, high viral titer and yield

Inactive Publication Date: 2011-11-16
长春生物制品研究所有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] The invention provides a method for producing rabies vaccine using a bioreactor and a sheet carrier to solve the problems of long production cycle, low virus yield and unsuitability for large-scale production in the current production of rabies vaccine

Method used

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  • Method for producing rabies vaccine by applying bioreactor and sheet carrier
  • Method for producing rabies vaccine by applying bioreactor and sheet carrier

Examples

Experimental program
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Effect test

Embodiment 1

[0039] (1) Sterilize the Celligen 310 14L bioreactor with a fixed bed basket stirring system and assemble it completely, add cell culture medium, add BioNOC II? cell culture disks microcarrier 15g / L, calf serum content in the culture medium is 5 %, adjust the pH to 7.0, temperature at 33°C, and stirring speed at 20rpm overnight; inoculate Vero cells amplified by 3L spinner bottles at a seeding density of 0.5×10 6 cells / ml, adjust CO 2 The pressure is 5%, the pH is 7.2, the temperature is 35°C, the dissolved oxygen is 20%, the glucose concentration is 25uM, and the stirring speed is 20rpm;

[0040] (2) When the cell density increases to 5×10 7 cells / ml was inoculated with aGV virus, the inoculation ratio was 1:100,

[0041] (3) Detect parameters such as glucose and lactic acid content through a biochemical analyzer, and adjust the reactor control parameters: CO 2 The pressure is 5%, the pH is 7.4, the temperature is 30°C, the dissolved oxygen is 20%, the glucose concentratio...

Embodiment 2

[0044] (1) Sterilize the Celligen 310 14L bioreactor with a fixed bed basket stirring system and assemble it completely, add cell culture medium, add BioNOC II? cell culture disks microcarrier 18g / L, calf serum content in the culture medium is 8 %, adjust the pH to 7.2, temperature 35°C, and stirring speed 35rpm overnight; inoculate Vero cells amplified by 3L spinner bottles, and the inoculation density is 1.3×10 6 cells / ml, adjust CO 2 The pressure is 8%, the pH is 7.3, the temperature is 36°C, the dissolved oxygen is 55%, the glucose concentration is 35uM, and the stirring speed is 60rpm;

[0045] (2) When the cell density increases to 5×10 7 cells / ml was inoculated with aGV virus, the inoculation ratio was 1:300,

[0046] (3) Detect parameters such as glucose and lactic acid content through a biochemical analyzer, and adjust the reactor control parameters: CO 2 The pressure is 8%, the pH is 7.6, the temperature is 33°C, the dissolved oxygen is 55%, the glucose concentrat...

Embodiment 3

[0049] (1) Sterilize the Celligen 310 14L bioreactor with a fixed bed basket stirring system and assemble it completely, add cell culture medium, add BioNOC II? cell culture disks microcarrier 20g / L, calf serum content in the culture medium is 10 %, adjust the pH to 7.4, temperature at 37°C, and stirring speed at 50rpm overnight; inoculate Vero cells amplified by 3L spinner bottles at a seeding density of 2×10 6 cells / ml, adjust CO 2 The pressure is 10%, the pH is 7.4, the temperature is 37°C, the dissolved oxygen is 90%, the glucose concentration is 40uM, and the stirring speed is 100rpm;

[0050] (2) When the cell density increases to 5×10 7 cells / ml was inoculated with aGV virus, the inoculation ratio was 1:500,

[0051] (3) Detect parameters such as glucose and lactic acid content through a biochemical analyzer, and adjust the reactor control parameters: CO 2 The pressure is 10%, the pH is 7.8, the temperature is 35°C, the dissolved oxygen is 90%, the glucose concentrat...

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Abstract

The invention relates to a method for producing rabies vaccines by applying a bioreactor and a sheet carrier, belonging to the technical field of biological products. The method comprises the following steps of: culturing cells by taking BioNOC IITM cell culture disks of the Taiwan SaiYu cell technology Co., Ltd as carriers in a Celligen 310 type 14 L bioreactor with a fixed bed basket-type stirring system; preparing rabies viruses with high titer through specific jar cell density, added viruses and the set parameters of the Celligen 310 type 14 L bioreactor; and producing the rabies vaccines. The method disclosed by the invention has the advantages of high density of cultured cells, high titer of the rabies viruses, high yield and low production cost; the Celligen 310 type 14 L bioreactor is enlarged in surface area used for cell adherence growth by adding the BioNOC IITM cell culture disks and then achieves the cell culture density at 5*108 / ml by creating good culture conditions; according to the method disclosed by the invention, the titer of the rabies viruses is correspondingly, greatly logarithmically enhanced and can maximally reach 1*109 FFU / ml, and therefore production cost is greatly reduced; in addition, the method disclosed by the invention is suitable for the large-scale production of rabies vaccines.

Description

technical field [0001] The invention relates to the technical field of biological products and is applied to the production of rabies vaccines. Background technique [0002] Rabies is a zoonotic disease caused by rabies virus, and humans and all warm-blooded animals are susceptible to rabies virus. Once the disease is 100% dead, the only effective way is to vaccinate effective human rabies vaccine. The currently used human rabies vaccines are human diploid cell vaccine (HDCV); primary hamster kidney cell human rabies vaccine (PHKCV); refined chicken embryo cell human rabies vaccine (PCECV); Vero cell human rabies vaccine ( PVRV); duck embryo human rabies vaccine (PDEV), etc. There are two types of human rabies vaccines produced in my country, PHKCV and PVRV. [0003] At present, the inactivated vaccines or live attenuated vaccines approved for production in my country, except for hepatitis B vaccine, are all prepared by the method of propagating viruses with mammalian cel...

Claims

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Application Information

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IPC IPC(8): A61K39/205C12N7/00A61P31/14C12R1/93
Inventor 郭秀侠杨屹苗丽刘岩蔡月红李宇辉崔文广姜文波井伟东苑志刚丁丽丽
Owner 长春生物制品研究所有限责任公司
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