文献类型：期刊文献

英文题名：High cell density perfusion process for high yield of influenza A virus production using MDCK suspension cells

作者：Wu, Yixiao[1,2];Bissinger, Thomas[2];Genzel, Yvonne[2];Liu, Xuping[1,3];Reichl, Udo[1,4];Tan, Wen-Song[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Max Planck Inst Dynam Complex Tech Syst, Bioproc Engn Grp, Sandtorstr 1, D-39106 Magdeburg, Germany;[3]Shanghai BioEngine Sci Tech Co Ltd, 781 Cailun Rd, Shanghai 201203, Peoples R China;[4]Otto von Guericke Univ, Chair Bioproc Engn, Univ Pl 2, D-39106 Magdeburg, Germany

年份：2021

卷号：105

期号：4

起止页码：1421

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20210509852366);WOS:【SCI-EXPANDED(收录号:WOS:000613036000001)】；

语种：英文

外文关键词：MDCK cells; Influenza A virus; ATF-based perfusion; Process optimization; Lab-scale bioreactors

摘要：Similar to the recent COVID-19 pandemic, influenza A virus poses a constant threat to the global community. For the treatment of flu disease, both antivirals and vaccines are available with vaccines the most effective and safest approach. In order to overcome limitations in egg-based vaccine manufacturing, cell culture-based processes have been established. While this production method avoids egg-associated risks in face of pandemics, process intensification using animal suspension cells in high cell density perfusion cultures should allow to further increase manufacturing capacities worldwide. In this work, we demonstrate the development of a perfusion process using Madin-Darby canine kidney (MDCK) suspension cells for influenza A (H1N1) virus production from scale-down shake flask cultivations to laboratory scale stirred tank bioreactors. Shake flask cultivations using semi-perfusion mode enabled high-yield virus harvests (4.25 log(10)(HAU/100 mu L)) from MDCK cells grown up to 41 x 10(6) cells/mL. Scale-up to bioreactors with an alternating tangential flow (ATF) perfusion system required optimization of pH control and implementation of a temperature shift during the infection phase. Use of a capacitance probe for on-line perfusion control allowed to minimize medium consumption. This contributed to a better process control and a more economical performance while maintaining a maximum virus titer of 4.37 log(10)(HAU/100 mu L) and an infectious virus titer of 1.83 x 10(10) virions/mL. Overall, this study clearly demonstrates recent advances in cell culture-based perfusion processes for next-generation high-yield influenza vaccine manufacturing for pandemic preparedness.