tetano
Editor, Senior Moderator
Appl Microbiol Biotechnol
. 2021 Jan 30;1-14.
doi: 10.1007/s00253-020-11050-8. Online ahead of print.
High cell density perfusion process for high yield of influenza A virus production using MDCK suspension cells
Yixiao Wu[SUP] 1 2 [/SUP], Thomas Bissinger[SUP] 2 [/SUP], Yvonne Genzel[SUP] 2 [/SUP], Xuping Liu[SUP] 3 4 [/SUP], Udo Reichl[SUP] 1 5 [/SUP], Wen-Song Tan[SUP] 1 [/SUP]
Affiliations
Abstract
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[SUB]10[/SUB](HAU/100 μL)) from MDCK cells grown up to 41 ? 10[SUP]6[/SUP] 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[SUB]10[/SUB](HAU/100 μL) and an infectious virus titer of 1.83 ? 10[SUP]10[/SUP] 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. KEY POINTS: • First MDCK suspension cell-based perfusion process for IAV produciton was established. • "Cell density effect" was overcome and process was intensified by reduction of medium use and automated process control. • The process achieved cell density over 40 ? 10[SUP]6[/SUP] cells/mL and virus yield over 4.37 log[SUB]10[/SUB](HAU/100 μL).
Keywords: ATF-based perfusion; Influenza A virus; Lab-scale bioreactors; MDCK cells; Process optimization.
. 2021 Jan 30;1-14.
doi: 10.1007/s00253-020-11050-8. Online ahead of print.
High cell density perfusion process for high yield of influenza A virus production using MDCK suspension cells
Yixiao Wu[SUP] 1 2 [/SUP], Thomas Bissinger[SUP] 2 [/SUP], Yvonne Genzel[SUP] 2 [/SUP], Xuping Liu[SUP] 3 4 [/SUP], Udo Reichl[SUP] 1 5 [/SUP], Wen-Song Tan[SUP] 1 [/SUP]
Affiliations
- PMID: 33515287
- PMCID: PMC7847233
- DOI: 10.1007/s00253-020-11050-8
Abstract
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[SUB]10[/SUB](HAU/100 μL)) from MDCK cells grown up to 41 ? 10[SUP]6[/SUP] 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[SUB]10[/SUB](HAU/100 μL) and an infectious virus titer of 1.83 ? 10[SUP]10[/SUP] 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. KEY POINTS: • First MDCK suspension cell-based perfusion process for IAV produciton was established. • "Cell density effect" was overcome and process was intensified by reduction of medium use and automated process control. • The process achieved cell density over 40 ? 10[SUP]6[/SUP] cells/mL and virus yield over 4.37 log[SUB]10[/SUB](HAU/100 μL).
Keywords: ATF-based perfusion; Influenza A virus; Lab-scale bioreactors; MDCK cells; Process optimization.