Production method of freeze dried chickenpox attenuated live vaccine and products therefrom

A live attenuated vaccine and production method technology, applied in the field of live attenuated varicella vaccine and products, can solve the problems of restricting the large-scale production of varicella vaccine, reducing the ability of VZV, and low VZV production, and achieve the removal or reduction of bovine serum residues Quantity, improve stability, reduce the effect of adverse reactions

Inactive Publication Date: 2005-03-16
CHANGCHUN KEYGEN BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of VZV in GPEC cells is very low
WI-38 and MRC-5 cell lines have high passage numbers, which reduces their ability to produce VZV
In addition, the static culture method is used in the process of cell and virus propagation in the above patent, so the above method limits the large-scale production of varicella vaccine

Method used

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  • Production method of freeze dried chickenpox attenuated live vaccine and products therefrom
  • Production method of freeze dried chickenpox attenuated live vaccine and products therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Take human embryonic lung diploid cell 2BS (32 generations) monolayer and subculture according to 1:4, adjust the rotating speed of the bottle machine to 30 rpm, culture at 37°C for 5 days, grow into a dense monolayer, and place in the culture bottle Add the virus seeds in the ratio of virus seeds to cells 1:60, infect the cells, and incubate at 35°C for 3 days until the cytopathic changes reach more than 75%, use Earle's solution three times the original maintenance solution volume Rinse the cell surface, wash away the bovine serum, and add the vaccine solution containing the vaccine stabilizer to harvest. The mass ratio of each component in the vaccine liquid is: 2.5% of sucrose, 2% of gelatin, 0.5% of sodium glutamate, 0.1% of urea, 0.1% of arginine, 0.3% of sorbitol and 1% of human serum albumin. Freeze and thaw the harvested vaccine solution at -70°C, break the cells by ultrasonic at 20KHz, centrifuge at 1000rpm at 4°C for 30 minutes to remove cell debris, clarify ...

Embodiment 2

[0055] Firstly, human embryonic lung diploid cells 2BS (passage 31) were subcultured at a ratio of 1:2. The medium used for cell expansion is MEM supplemented with 10% calf serum, pH 7.2. Place the cell culture flask at 37°C. Culture under rotation for 3 days (20 rpm / hour) to form a uniform and dense monolayer of cells, and then discard the growth solution. Virus seeds were added to the culture flask at a ratio of 1:80 virus seeds to cells for infection, and cultured in rotation at 35° C. (rotating speed 20 rpm) for 4 days. When more than 75% of the typical lesions appear on the 2BS cells, pour out the original maintenance solution, wash the cell surface with Earle's solution four times the original maintenance solution volume, wash away the bovine serum, and add the vaccine solution containing the vaccine stabilizer to harvest. The mass ratio of each component in the vaccine liquid is: 2.5% of sucrose, 2% of gelatin, 0.5% of sodium glutamate, 0.1% of urea, 0.1% of arginine,...

Embodiment 3

[0057] Firstly, human embryonic lung diploid cells 2BS (passage 38) were subcultured at a ratio of 1:2. The medium used for cell expansion is MEM supplemented with 12% calf serum, pH 7.2. Place the cell culture bottle at 37° C. for 4 days under rotation (25 rpm / hour) to form a uniform and dense monolayer of cells, and then discard the growth medium. Virus seeds were added to the culture flask at a ratio of 1:90 virus seeds to cells for infection, and rotated at 35° C. (15 revolutions / hour) for 4 days. When more than 75% of the typical lesions appear on the 2BS cells, pour out the original maintenance solution, wash the cell surface with Earle's solution three times the original maintenance solution volume, wash away the bovine serum, and add the vaccine solution containing the vaccine stabilizer to harvest. The mass ratio of each component in the vaccine liquid is: 1.5% of sucrose, 4% of gelatin, 0.5% of sodium glutamate, 0.2% of urea, 0.1% of arginine, 0.4% of sorbitol and 0...

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Abstract

The invention provides the production method of freeze dried chickenpox attenuated live vaccine and products made thereby, wherein the method consists of using chickenpox attenuation Oka as strain, employing rotary bottle manufacturing process, and using diploid cell 2BS as culture medium mass. The invention can realize stabilized vaccine virus droplet degree and increased single bottle output compared with the static culture method.

Description

Technical field: [0001] The invention relates to a live attenuated varicella vaccine and its products, and in particular discloses a production method of a freeze-dried live attenuated varicella vaccine. The attenuated strain of varicella virus (Oka strain) is used to infect human embryonic lung diploid cells (2BS ) The method for preparing live attenuated varicella vaccine by rotary culture belongs to the technical field of vaccine production and preparation. technical background: [0002] The production of viral vaccines usually uses chicken embryo cells, brain cells of young mice and diploid cells of mammals as host cells. But they do not have advantages due to the low sensitivity of the virus, the complicated purification process and the side effects due to the presence of foreign protein contamination. On the contrary, the use of normal diploid cells of human origin can reduce the side effects caused by foreign proteins, so they are more preferable in the preparation o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/19A61K39/25A61K47/42A61P31/20
Inventor 王连成刘晔李占春王鹏哈秀杰李延成麻广李春明李生军王瑛杰李开军徐俊峰王丽华宣黎波刘延威王斐戴长河杨倩
Owner CHANGCHUN KEYGEN BIOLOGICAL PROD
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