Performance of small-scale CHO perfusion cultures using an acoustic cell filtration device for cell retention: Characterization of separation efficiency and impact of perfusion on product quality
Article first published online: 16 JUN 2000
Copyright © 2000 John Wiley & Sons, Inc.
Biotechnology and Bioengineering
Volume 69, Issue 4, pages 440–449, 20 August 2000
How to Cite
Ryll, T., Dutina, G., Reyes, A., Gunson, J., Krummen, L. and Etcheverry, T. (2000), Performance of small-scale CHO perfusion cultures using an acoustic cell filtration device for cell retention: Characterization of separation efficiency and impact of perfusion on product quality. Biotechnol. Bioeng., 69: 440–449. doi: 10.1002/1097-0290(20000820)69:4<440::AID-BIT10>3.0.CO;2-0
- Issue published online: 16 JUN 2000
- Article first published online: 16 JUN 2000
- Manuscript Accepted: 30 MAR 2000
- Manuscript Received: 14 JUL 1999
- acoustic filtration;
- separation efficiency;
- Chinese hamster ovary (CHO) cell culture;
- product quality;
Several small-scale Chinese hamster ovary (CHO) suspension cultures were grown in perfusion mode using a new acoustic filtration system. The separation performance was evaluated at different cell concentrations and perfusion rates for two different CHO cell lines. It was found that the separation performance depends inversely on the cell concentration and perfusion rate. High media flow rates as well as high cell concentrations resulted in a significant drop in the separation performance, which limited the maximal cell concentration achievable. However, packed cell volumes of 10% to 16% (corresponding to 3 to 6 · 107 cells/mL) could be reached and were maintained without additional bleeding after shifting the temperature to 33°C. Perfusion, up to 50 days, did not harm the cells and did not result in a loss of performance of the acoustic filter as often seen with other perfusion systems. Volumetric productivities in perfusion mode were 2- to 12-fold higher for two cell lines producing two different glycoproteins when compared to fed-batch or batch processes using the same cell lines. Product concentrations were in the range of 20% to 80% of batch or fed-batch culture, respectively. In addition, using the protease-sensitive product rhesus thrombopoietin, we could show that cultivation in perfusion mode drastically reduced proteolysis when compared to a batch culture without addition of protease inhibitors such as leupeptin. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 69: 440–449, 2000.