Strategy for targeted delivery of key nutrients in high cell density perfusion
October 6-10, 2019
High cell density perfusion is an attractive way for process intensification. We have developed high cell density processes using CHO cells for the production of monoclonal antibody, or using HEK293 cells for the production non-antibody glycoproteins.
A key aspect in the development of high cell density process is to obtain low culture medium renewal, in order to decrease the medium usage, the volume of harvest and the logistics burden. The concentration of culture medium is one of the tools available to achieve this. We have obtained interesting results of very low cell specific perfusion rate (CSPR) down to one reactor volume medium renewed per day sustaining healthy culture at 100 x 106 cells/mL. The limitation of such an approach is that all the medium components are concentrated generating media close to the precipitation. Beyond the concentration of the whole medium formulation, it is attractive to have a method to deliver selected components in the culture.
Such a method was developed and successfully applied for the delivery of glucose to reduce the lactate production, which drastically impeded the cell growth, in HEK293 cell culture at 80 to 100 x 106 cells/mL for the production of erythropoietin. It was then systematically adopted for all the perfusion cultures in our lab, i.e. using CHO cells or HEK293 cells. The method was then applied to deliver sugars different from glucose in antibody producing CHO cell perfusion process in a study of the effect of different sugars on the glycosylation. Finally, it was applied for the delivery of the amino acids in high density CHO cell perfusion process for the production of antibody at very low CSPR.
Veronique Chotteau, Hubert Schwarz, Liang Zhang, Ye Zhang, Caijuan Zhan, Matthew Cheeks, Richard Turner, Andreas Castan, and Francisco Vilaplana, "Strategy for targeted delivery of key nutrients in high cell density perfusion" in "Integrated Continuous Biomanufacturing IV", Veena Warikoo, Roche, USA Alois Jungbauer, BOKU, Austria Jon Coffman, Boehringer Ingelheim, USA Jason Walther, Sanofi, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/biomanufact_iv/60