Method for industrially producing animal influenza vaccine by using bioreactor
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
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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);
[0026] Virus: H9N2 subtype avian influenza virus ZB1 strain;
[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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