Method for industrially producing animal influenza vaccine by using bioreactor
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A bioreactor, influenza vaccine technology, applied in the field of veterinary vaccine production, achieves the effects of high cell density, stable and reliable quality, and low labor intensity
Inactive Publication Date: 2011-07-27
WUHAN CHOPPER BIOLOGY
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However, there are no reports on the application of specific bioreactor technology to high-density culture of host cells through step-by-ste...
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Embodiment 1
[0024] Bioreactor: 14L ~ 40L bioreactor of NBS Company in the United States;
[0025] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);
[0027] Cell growth medium: DMEM medium containing 8% serum by volume (Beijing Qingda Tianyi Biotechnology Co., Ltd.);
[0028] Cell maintenance medium: DMEM medium containing 1 μg / ml trypsin (Beijing Qingda Tianyi Biotechnology Co., Ltd.);
[0029] Cell culture: In 14L bioreactors, add Cytodex-1 at a concentration of 10g / L, after hydration, wash with pH7.2 phosphate buffer solution PBS several times, add cell growth solution after sterilization, and inoculate MDCK cells were cultured. The parameters of the culture method are: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm; samples are taken regularly every day to observe the growth of the cells, count the cells, and measure the consumption of glucose. When the density of th...
Embodiment 2
[0034] Bioreactor: 14L ~ 40L bioreactor of NBS company in the United States
[0035] Microcarrier: Cytodex-1 (GE Life Sciences, please write the full name)
[0036]Virus: Swine influenza virus H1N1 subtype
[0037] Cell growth medium: M199 medium containing 8% serum
[0038] Cell maintenance medium: M199 medium containing 1 μg / ml trypsin
[0039] Cell culture: In 14L bioreactors, add Cytodex-1 at a concentration of 10g / L, after hydration, wash with pH7.2 phosphate buffer solution PBS several times, add cell growth solution after sterilization, and inoculate Vero cells were cultured. The parameters of the culture method are: pH 7.2, temperature 37°C, dissolved oxygen 50%, stirring speed 30-100rpm; samples are taken regularly every day to observe the growth of the cells, count the cells, and measure the consumption of glucose. When the density of the cells reaches 1.5×10 6 Perfusion starts at 0.5 / ml, and the perfusion speed depends on the density of cells and the consumption...
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Abstract
The invention relates to a method for industrially producing animal influenza vaccine by using a bioreactor, comprising the following steps of: (1) adding Vero or MDCK cells in the sterilized bioreactor containing a micro-carrier and culturing in a serum-including culture medium; (2) replacing the culture medium including serum in the bioreactor by a serum-free culture medium which includes pancreatin; (3) inoculating animal influenza virus to culture continuously; (4) harvesting to obtain a cell-cultured influenza virus stock solution when the pathological change of the cells is up to 80%; and (5) hyper-filtering, concentrating and inactivating the influenza virus stock solution and then adding an adjuvant to obtain the influenza inactivated vaccine. The method provided by the invention has the advantages of controllable process parameters, high cell density, high virus titer, high production efficiency, low labor intensity and the like. The influenza vaccine produced by the invention has good security, stable and reliable quality, small side reaction and high immune effect and has a better immune protection function for the attack of the influenza virus.
Description
technical field [0001] The invention relates to the production of veterinary vaccines, in particular to a method for industrially producing animal influenza inactivated vaccines using a bioreactor. Background technique [0002] Influenza viruses belong to the Orthomyxoviridae family of RNA viruses and are divided into three types: A, B, and C. Among them, influenza A virus is mainly found in poultry, and some subtypes can also infect various mammals such as pigs, horses, seals and whales, as well as humans; influenza B and C viruses are found in seals and pigs, respectively. Avian Influenza (AI) is a highly contagious infectious disease of poultry and wild poultry caused by type A influenza virus. Wild birds are the natural host of avian influenza virus. It has been confirmed that avian influenza virus can directly infect humans from poultry, causing human morbidity and death. Swine influenza (SI) is an acute, febrile and highly contagious respiratory infectious disease of...
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