Identification of process conditions influencing protein aggregation in Chinese hamster ovary cell culture

Citations Over TimeTop 25% of 2017 papers

Abstract

Protein aggregation of monoclonal antibodies (mAbs) is a common phenomenon associated with the production of these biopharmaceuticals. These aggregates can lead to adverse side effects in patients upon administration, thus expensive downstream processing steps to remove the higher molecular weight species are inevitable. A preferable approach is to reduce the level of aggregation during bioprocessing by a careful adjustment of critical process parameters. Recently, new analytical methods enabled characterization of mAb aggregation during bioprocessing of mammalian cells. Furthermore, rapid and efficient bioprocess optimization has been performed using design of experiments (DoE) strategies. In this work, we describe a DoE-based approach for the analysis of process parameters and cell culture additives influencing protein aggregation in Chinese hamster ovary (CHO) cell cultures. Important bioprocess variables influencing the aggregation of mAb and host cell proteins were identified in initial screening experiments. Response surface modeling was further applied in order to find optimal conditions for the reduction of protein aggregation during cell culture. It turned out that a temperature-shift to 31 °C, osmolality above 420 mOsm/kg, agitation at 100 rpm and 0.04% (w/v) antifoam significantly reduced the level of aggregates without substantial detrimental effects on cell culture performance in our model system. Finally, the aggregation reducing conditions were verified and applied to another production system using a different bioprocess medium and another CHO cell line producing another mAb. Our results show that protein aggregation can be controlled during cell culture and helps to improve bioprocessing of mAbs, by giving insights into the protein aggregation at its origin in mammalian cell culture.

Related Papers

• → Upstream and Downstream Bioprocessing in Enzyme Technology(2023)25 cited
• → Production of monoclonal antibodies in COS and CHO cells(1995)115 cited
• → Identification of process conditions influencing protein aggregation in Chinese hamster ovary cell culture(2017)29 cited
• → A novel approach to characterize host cell proteins associated with therapeutic monoclonal antibodies(2017)12 cited
• → Computer Aided Design of Large Scale Integrated Bioprocess(1998)