Method for producing avian influenza vaccine by using cells cultured by spherical microcarrier bioreactor

A bioreactor and cell culture technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as limited production output, and achieve the effect of shortening the production cycle

Inactive Publication Date: 2012-09-12
ZHAOQING DAHUANONG BIOLOGIC PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the amount of virus produced only by animal cell culture is limited, so the application of vector culture technology to the production of av...

Method used

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  • Method for producing avian influenza vaccine by using cells cultured by spherical microcarrier bioreactor
  • Method for producing avian influenza vaccine by using cells cultured by spherical microcarrier bioreactor
  • Method for producing avian influenza vaccine by using cells cultured by spherical microcarrier bioreactor

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Experimental program
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Embodiment 1

[0026] Embodiment 1: microcarrier bioreactor suspension culture breeding H5 subtype avian influenza inactivated vaccine

[0027] 1: Select the type of cell MDCK required for the introduction of the vector;

[0028] 2: The method of spinner bottle propagating cell passage: first resuscitate the seed cell MDCK selected in step 1 into a T225 cell flask, grow for about 48 hours, and passage according to the ratio of 1:4; then put 4 full-celled T225 cells The cells in the bottle were transferred to a 15L spinner bottle for culture; the cells were subcultured at a ratio of 1:3 for about 48 hours; the medium used in this process was MEM, the serum was fetal bovine serum, and the amount used was 7%;

[0029] 3: Harvest the cells in the spinner bottle: wash the cells obtained in step 2 with PBS 2-3 times, then add 125ml of 0.25% trypsin-EDTA solution to digest and collect the cells;

[0030] 4: After counting the harvested cells, the selected microcarrier is Cytodex I, and the cells a...

Embodiment 2

[0046] Embodiment 2: Microcarrier suspension culture propagation H9 subtype avian influenza virus

[0047] 1: Select the type of cell MDCK required for the introduction of the vector;

[0048] 2: The method of spinner bottle propagating cell passage: first resuscitate the seed cell MDCK selected in step 1 into a T225 cell flask, grow for about 48 hours, and passage according to the ratio of 1:4; then put 4 full-celled T225 cells The cells in the bottle were transferred to a 15L spinner bottle for culture; the cells were subcultured at a ratio of 1:4 for about 48 hours; the medium used in this process was MEM, the serum was fetal bovine serum, and the amount used was 7-10 %;

[0049] 3: Harvest the cells in the spinner bottle: wash the cells obtained in step 2 with PBS 2-3 times, then add 125ml of 0.25% trypsin-EDTA solution to digest and collect the cells;

[0050] 4: After the harvested cells are counted, the selected microcarrier is hyclone, and the cells are inoculated ac...

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Abstract

The invention provides a method for producing an avian influenza vaccine by using cells cultured by a spherical microcarrier bioreactor. The method comprises the following steps of: selecting seed cells; passaging cells reproduced in a spinner bottle; harvesting the cells in the spinner bottle; inoculating the cells; culturing the inoculated cells in the bioreactor; inoculating avian influenza virus; reproducing the avian influenza virus; collecting avian influenza virus liquid; inactivating the avian influenza virus liquid; preparing an avian influenza cell vaccine; and immunizing the avian influenza cell vaccine. By the method, the defects that the avian influenza vaccine produced by the traditional process is long in production period, large in inter-batch difference, high in endotoxin content and influenced by the supply of embryo eggs, and a tablet carrier is difficult to amplify are overcome; and the prepared avian influenza vaccine product is small in inter-batch difference, free from bacterial pollution, low in endotoxin content and formaldehyde content, stable in quality and short in the production period, and the tablet carrier is easy to amplify.

Description

technical field [0001] The invention relates to the technical field of bird flu vaccines, in particular to a method for producing bird flu vaccines by using spherical microcarrier bioreactors to cultivate cells. Background technique [0002] In the past, our country has been producing influenza vaccines through chicken embryo culture. Since the production of influenza vaccine from chicken eggs requires the consumption of a large number of chicken embryos, the production cycle is long, it is easy to be polluted, and it is not easy to control, and each fertilized egg can only be injected with one virus strain at a time, so this production method is very inefficient and is not conducive to large-scale flu outbreak. [0003] Using mammalian cells as the matrix to prepare vaccines has the advantages of no exogenous factor contamination, easy large-scale production, and better stability of virus antigens. However, the amount of virus produced only by animal cell culture is limit...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61P31/14C12N7/00C12N7/06C12R1/93
Inventor 陈瑞爱徐家华施维松张冬霞汤钦
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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