Monitoring, Design and Optimization of Bioprocesses

Header image 1377178557

Monitoring, Design and Optimization of Bioprocesses

With new cell substrates available for vaccine production additional process strategies have become an option. Today, if possible, suspension cells are used that grow in chemically defined media. They can be used for more than 100 passages and grow in batch culture easily to 5-10 x 106 cells/mL.

For suspension CHO cells to produce monoclonal antibodies perfusion and fed-batch strategies are today state-of-the-art, allowing processes to run for several weeks leading to continuous upstream and downstream approaches with high product yields. Good perfusion media or special feeds have been developed.

However, for the cell lines used in viral vaccine manufacturing these media are not readily available. Typically, at higher cell densities (> 4 x 106 cells/mL) a “high cell density effect” leads to loss of cell-specific productivity. Therefore, the question is, if these new substrates allow overcoming this high cell density effect for some viruses. If yes, what cell densities are possible (maximum CHO cell density described: 5 x 108 cells/mL)? Additionally it is addressed, what alternative process options, especially towards disposable bioreactors and continuous processes are possible for vaccine manufacturing today.

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)
 
Go to Editor View
loading content