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Preparation method for measles attenuated live vaccine

A technology of live attenuated vaccine and measles virus, applied in the field of vaccines, can solve the problems of easy inactivation of measles virus, poor uniformity of vaccine quality, and ineffectiveness of vaccines, and achieve easy large-scale production, increase single batch yield, and uniform quality and stable effect

Inactive Publication Date: 2012-04-11
CHENGDU KANGHUA BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, although the current measles vaccine has some control over the bovine serum albumin content of the virus harvest in terms of production, it does not strictly control the residues of chicken embryo cell components. These heterologous proteins are caused by side reactions during vaccination. important factor, and the titer of the virus culture in the conventional production process itself is not high, and the measles virus is easily inactivated in a liquid environment. If purification and other processes are used, the virus titer will not meet the production requirements and the vaccine will fail
At the same time, the current measles vaccine is produced by a bottle-type process, the production batch is small, the quality uniformity of the produced vaccine is poor, and pollution is prone to occur due to complicated operations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A preparation method of live attenuated measles vaccine, comprising the following steps:

[0020] Step 1: Microcarrier bioreactor culture: mix chicken embryo cells and measles virus strains and inoculate them into a microcarrier bioreactor, cultivate the mixture at a temperature of 33°C and a pH of 7.2, replace the nutrient solution every day, and cultivate to a cell concentration of 10 6 cells / ml, the chicken embryo cells were taken from SPF chicken embryos incubated at the age of 9 days, and the concentration was 1×10 5 / ml; described measles virus strain is Schwarz strain, and its inoculum size is 0.001MOI; The microcarrier in described microcarrier bioreactor is spherical microcarrier, and its concentration is 2g / L;

[0021] Step 2: Maintenance medium culture: observe the cells every day, when the chicken embryo cells begin to have specific lesions, wash the culture several times, and replace the serum-free MEM maintenance medium to culture the virus;

[0022] Ste...

Embodiment 2

[0028] A preparation method of live attenuated measles vaccine, comprising the following steps:

[0029] Step 1: microcarrier bioreactor culture: mix chicken embryo cells and measles virus strains and inoculate them into a microcarrier bioreactor, cultivate the mixture at a temperature of 33°C and a pH of 7.6, and replace the cell growth medium every day. Cultivate until the cell concentration reaches 10 7 cells / ml, the chicken embryo cells were taken from SPF chicken embryos incubated for 10 days, and the concentration was 5×10 5 / ml; the measles virus strain is a Moraten strain, and its inoculum size is 0.005MOI; the microcarrier in the microcarrier bioreactor is a porous spherical microcarrier, and its concentration is 10g / L;

[0030] Step 2: Maintenance medium culture: observe the cells every day, when the chicken embryo cells begin to have specific lesions, wash the culture several times, and replace the serum-free MEM maintenance medium to culture the virus;

[0031] S...

Embodiment 3

[0037] A preparation method of live attenuated measles vaccine, comprising the following steps:

[0038] Step 1: Microcarrier bioreactor culture: Mix chicken embryo cells and measles virus strains and inoculate them into microcarrier bioreactors, cultivate the mixture at a temperature of 33°C and a pH of 7.5, replace the nutrient solution every day, and cultivate to a cell concentration of 10 6 cells / ml, the chicken embryo cells were taken from SPF chicken embryos incubated at the age of 9 days, and the concentration was 3×10 5 / ml; described measles virus strain is Schwarz strain, and its inoculum size is 0.003MOI; The microcarrier in described microcarrier bioreactor is a fiber structure carrier, and its concentration is 6g / L;

[0039] Step 2: Maintenance medium culture: observe the cells every day, when the chicken embryo cells start to have specific lesions, wash the culture multiple times, and replace the serum-free maintenance medium to culture the virus;

[0040] Step...

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Abstract

The present invention relates to the field of vaccine, especially to a preparation method for a measles attenuated live vaccine. The method comprises the following steps: culturing in a microcarrier bioreactor; culturing by a maintenance media; harvesting and purifying; preparing the vaccine and freeze drying. With adopting the high density culture method of the bioreactor, it is ensured that the resulting virus titer is higher than the virus titer from the conventional process; on the premise of the high virus titer by the method of the present invention, a molecular sieve chromatography method or a hollow fiber filtering method is further adopted to carry out purification to remove most of impurity components, such that the generation of the side reactions of the vaccination is effectively avoided. In addition, with adopting the bioreactor to culture the virus, the culture conditions are stable, such that the large-scale production is easy to realize, and the single batch production of the vaccine is increased so as to ensure the quality uniformity and the quality stability of the product.

Description

technical field [0001] The invention relates to the field of vaccines, in particular to a preparation method of live attenuated measles vaccine. Background technique [0002] Measles is an ancient infectious disease caused by infection with the measles virus. At present, vaccination with live attenuated measles vaccine is an effective means to prevent and control the occurrence of the disease. The effect of live attenuated measles vaccine is mainly based on the survival and reproduction of live virus in the body. Because measles virus is very sensitive to light and heat, it is easy to be inactivated, and its stability is related to the temperature of the virus environment and the protein content in the virus fluid. It is not easy to be inactivated during the process and is prepared into a freeze-dried dosage form. [0003] However, although the current measles vaccine has some control over the bovine serum albumin content of the virus harvest in terms of production, it do...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/165A61P31/14
Inventor 赵志鹏侯文礼
Owner CHENGDU KANGHUA BIOLOGICAL PROD
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