Insights into large-scale cell-culture reactors: II. Gas-phase mixing and CO₂ stripping.

Most discussions about stirred tank bioreactors for cell cultures focus on liquid-phase motions and neglect the importance of the gas phase for mixing, power input and especially CO(2) stripping. Particularly in large production reactors, CO(2) removal from the culture is known to be a major problem. Here, we show that stripping is mainly affected by the change of the gas composition during the movement of the gas phase through the bioreactor from the sparger system towards the headspace. A mathematical model for CO(2)-stripping and O(2)-mass transfer is presented taking gas-residence times into account. The gas phase is not moving through the reactor in form of a plug flow as often assumed. The model is validated by measurement data. Further measurement results are presented that show how the gas is partly recirculated by the impellers, thus increasing the gas-residence time. The gas-residence times can be measured easily with stimulus-response techniques. The results offer further insights on the gas-residence time distributions in stirred tank reactors.

[1]  W. Miller,et al.  Effects of CO2 and osmolality on hybridoma cells: growth, metabolism and monoclonal antibody production , 1998, Cytotechnology.

[2]  A. Nienow Reactor Engineering in Large Scale Animal Cell Culture , 2006, Cytotechnology.

[3]  J M Smith,et al.  Development of a strategy to control the dissolved concentrations of oxygen and carbon dioxide at constant shear in a plant cell bioreactor , 1990, Biotechnology and bioengineering.

[4]  Marco Jenzsch,et al.  Insights into large-scale cell-culture reactors: I. Liquid mixing and oxygen supply. , 2011, Biotechnology journal.

[5]  R. Higbie,et al.  The Rate of Absorption of a Pure Gas into a Still Liquid during Short Periods of Exposure , 1935 .

[6]  D K Robinson,et al.  Industrial choices for protein production by large-scale cell culture. , 2001, Current opinion in biotechnology.

[7]  Zizhuo Xing,et al.  Scale‐up analysis for a CHO cell culture process in large‐scale bioreactors , 2009, Biotechnology and bioengineering.

[8]  D. Inlow,et al.  CO2 in large-scale and high-density CHO cell perfusion culture , 2004, Cytotechnology.

[9]  P N Royce,et al.  Effect of changes in the pH and carbon dioxide evolution rate on the measured respiratory quotient of fermentations , 1992, Biotechnology and bioengineering.

[10]  Weichang Zhou,et al.  NS0 cell damage by high gas velocity sparging in protein‐free and cholesterol‐free cultures , 2008, Biotechnology and bioengineering.

[11]  Nina F. Thornhill,et al.  Estimation of dissolved carbon dioxide concentrations in aerobic fermentations , 1991 .

[12]  A. Lübbert,et al.  Catchment area of the cavities behind a rushton turbine in gassed-stirred-tank reactors , 1997 .

[13]  Sigma S Mostafa,et al.  Strategies for Improved dCO2 Removal in Large‐Scale Fed‐Batch Cultures , 2003, Biotechnology progress.

[14]  A. Nienow,et al.  Measurement of gas and liquid flows in stirred tank reactors with multiple agitators , 2009 .

[15]  A Lübbert,et al.  Fluid dynamics in bubble column bioreactors: experiments and numerical simulations. , 2000, Biotechnology and bioengineering.