Method for proliferating avian influenza viruses in bioreactor with cell carrier

An avian influenza virus and bioreactor technology, which is applied in the biological field, can solve problems such as large-scale vaccine production of unseen avian influenza viruses, and achieve the effects of improving purity and safety, realizing automatic control and uniform products.

Inactive Publication Date: 2011-07-20
深圳市南海潮生物技术有限公司
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Problems solved by technology

[0005] my country's biological products industry uses more primary animal cells in the production of various vaccines, while passage cells are ra

Method used

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  • Method for proliferating avian influenza viruses in bioreactor with cell carrier
  • Method for proliferating avian influenza viruses in bioreactor with cell carrier
  • Method for proliferating avian influenza viruses in bioreactor with cell carrier

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Embodiment

[0022] Reagents used: MDCK cells were purchased from ATCC library, and microcarrier Cytodex1 was purchased from Pharmacia Biotech AB Uppsala Sweden (batch number: 223429).

[0023] After swelling per gram of Cytodex1 (dry powder), the surface area is 4400cm 2 , the amount of microcarriers is 5-10g / L. Cell density in the reactor=4400×10×166666 / 1000=7.3×106 cells / ml. The density of inoculated MDCK cells was 6.6×106 cells / ml. The H9N2 avian influenza virus was diluted 1:1000 times with DMEN medium containing 0.025 concentration of trypsin and inoculated into the cells. After 1 hour of adsorption, it was cultured in serum-free DMEN medium. Harvest the virus, harvest the virus liquid with 50% of the liquid exchange volume, harvest the virus liquid completely in 48 hours, and stop the bioreactor cultivation. Culture parameters: Stirring speed 40 rev / min: Dissolved oxygen reaches ±50% during the cell growth period, and ±40% during the cell expression period. Ventilation: The vent...

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Abstract

The invention discloses a method for replicating and proliferating avian influenza viruses in the passage cells which are absorbed and grow in the microcarrier of a bioreactor. The invention is based on the method which uses cells as carrier to proliferate viruses in the bioreactor. The obtained avian influenza viruses and the avian influenza virus vaccine product have uniform quality and stable toxicity. The automatic control of vaccine production can be realized, the problem of large-scale production and application can be solved, the production matrix of the influenza vaccine can be completely changed, the technology that chick embryo is adopted to culture viruses can be changed, the problems caused by chick embryo such as adventitious agent pollution and viral pollution and the interferences caused by heterologous proteins such as chick embryo can be reduced, the purity, safety and immune effect of vaccine can be increased and the method can play an important role in the prevention of avian influenza.

Description

technical field [0001] The invention belongs to the field of biology, in particular to a process for mass-propagating viruses in a bioreactor by cultivating cells with microcarriers. Background technique [0002] my country has a vast source of wealth, agriculture accounts for 70% of my country's total output value, and aquaculture is an important part of my country's farmers' economic sources. With the occurrence of avian influenza and the large-scale death of poultry, the production of preventive vaccines in my country is far from meeting the needs of the existing aquaculture market, and some vaccines are not effective, which aggravates the plight of the vaccine industry . Therefore, intensifying efforts to develop vaccines is the need of the current epidemic situation and the need of developing our country's national economy. [0003] At present, the virus sources of avian influenza vaccines all come from the culture of chicken embryos. The method is: inoculate the influ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/145C12R1/93
Inventor 徐安龙任政华王磊元少春张仁伟周亮潘武广周茂军吴赟覃梦颖陈枚花强媛媛王菲菲冯宇超
Owner 深圳市南海潮生物技术有限公司
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