Monitoring, Design and Optimization of Bioprocesses

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Monitoring, Design and Optimization of Bioprocesses

With new cell lines designed for vaccine production, various options for establishment of advanced process strategies are available. For ease of scale-up, suspension cell lines are used, whenever available, that grow in chemically defined media. Ideally, these cells can be used for more than 100 passages and grow in batch culture easily to 5-10 x 106 cells/mL.

For recombinant protein production, perfusion and fed-batch strategies using suspension CHO cells are state-of-the-art. The corresponding processes are running over several weeks leading to (semi-)continuous manufacturing with high product yields. To further support process intensification, optimized perfusion media and special feeding schemes have been developed.

For the cell lines used in viral vaccine manufacturing, however, such media are not readily available. Typically, at cell densities exceeding 4 x 106 cells/mL), the so-called “high cell density effect” often leads to a significant drop in cell-specific yield. Accordingly, better substrate formulations and process strategies need to be developed and tested comprehensively to overcome this crucial limitation. Additionally, alternative process options, especially towards disposable, single-use bioreactors and fully continuous processes need to be explored for future high-yield vaccine manufacturing.

Scale-up and different cultivation vessels

<i>Static growth in T-flasks and roller bottles</i> Zoom Image
Static growth in T-flasks and roller bottles
<i>Microcarrier systems in spinner flasks</i> Zoom Image
Microcarrier systems in spinner flasks
<i>5-15 L bioreactor</i> Zoom Image
5-15 L bioreactor
<i>2-10 L wave bioreactor</i> Zoom Image
2-10 L wave bioreactor

References

1.
Gallo Ramirez, L. E.; Nikolay, A.; Genzel, Y.; Reichl, U.: Bioreactor concepts for cell culture-based viral vaccine production. Expert Review of Vaccines 14 (9), pp. 1181 - 1191 (2015)
2.
Genzel, Y.: Designing cell lines for viral vaccine production: Where do we stand? Biotechnology Journal 10 (5), pp. 728 - 740 (2015)
3.
Genzel, Y.; Vogel, T.; Buck, J.; Behrendt, I.; Vazquez-Ramirez, D.; Schiedner, G.; Jordan, I.; Reichl, U.: High cell density cultivations by alternating tangential flow (ATF) perfusion for influenza A virus production using suspension cells. Vaccine 32 (24), pp. 2770 - 2781 (2014)
4.
Genzel, Y.; Rödig, J.; Rapp, E.; Reichl, U.: Vaccine production: Upstream processing with adherent or suspension cell lines. In: Animal Cell Biotechnology: Methods and Protocols, pp. 371 - 393 (Ed. Pörtner, R.). HUMANA Press Inc., New York (2014)
5.
Tapia, F.; Vogel, T.; Genzel, Y.; Behrendt, I.; Hirschel, M.; Gangemi, J. D.; Reichl, U.: Production of high-titer human influenza A virus with adherent and suspension MDCK cells cultured in a single-use hollow fiber bioreactor. Vaccine 32 (8), pp. 1003 - 1011 (2014)
6.
Peschel, B.; Frentzel, S.; Laske, T.; Genzel, Y.; Reichl, U.: Comparison of influenza virus yields and apoptosis-induction in an adherent and a suspension MDCK cell line. Vaccine 31 (48), pp. 5693 - 5699 (2013)
7.
Frensing, T.; Heldt, S.; Pflugmacher, A.; Behrendt, I.; Jordan, I.; Flockerzi, D.; Genzel, Y.; Reichl, U.: Continuous Influenza Virus Production in Cell Culture Shows a Periodic Accumulation of Defective Interfering Particles. PLoS One 8 (9), p. e72288 (2013)
 
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