Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
Appl Microbiol Biotechnol. 2013 Feb;97(3):1063-70. doi: 10.1007/s00253-012-4375-7. Epub 2012 Sep 4.
A process for human influenza H1N1 virus vaccine production from Madin-Darby canine kidney (MDCK) cells using a novel packed-bed bioreactor is described in this report. The mini-bioreactor was used to study the relationship between cell density and glucose consumption rate and to optimize the infection parameters of the influenza H1N1 virus (A/New Caledonia/20/99). The MDCK cell culture and virus infection were then monitored in a disposable perfusion bioreactor (AmProtein Current Perfusion Bioreactor) with proportional-integral-derivative control of pH, dissolved O(2) (DO), agitation, and temperature. During 6 days of culture, the total cell number increased from 2.0 × 10(9) to 3.2 × 10(10) cells. The maximum virus titers of 768 hemagglutinin units/100 μL and 7.8 × 10(7) 50 % tissue culture infectious doses/mL were obtained 3 days after infection. These results demonstrate that using a disposable perfusion bioreactor for large-scale cultivation of MDCK cells, which allows for the control of DO, pH, and other conditions, is a convenient and stable platform for industrial-scale production of influenza vaccines.
本报告介绍了一种使用新型填充床生物反应器从 Madin-Darby 犬肾 (MDCK) 细胞生产人季节性流感 H1N1 病毒疫苗的方法。该迷你生物反应器用于研究细胞密度与葡萄糖消耗率之间的关系，并优化流感 H1N1 病毒（A/New Caledonia/20/99）的感染参数。然后，在具有 pH、溶解氧 (DO)、搅拌和温度比例积分微分控制的一次性灌注生物反应器 (AmProtein Current Perfusion Bioreactor) 中监测 MDCK 细胞培养和病毒感染。在 6 天的培养过程中，总细胞数从 2.0×10(9)增加到 3.2×10(10)个细胞。感染后 3 天，获得了 768 血凝素单位/100μL 和 7.8×10(7)50%组织培养感染剂量/mL 的最大病毒滴度。这些结果表明，使用可控制 DO、pH 和其他条件的一次性灌注生物反应器进行 MDCK 细胞的大规模培养是流感疫苗工业规模生产的便捷、稳定的平台。
