A novel clarification approach for intensified monoclonal antibody processes with 100 million cells/mL using a single-use fluidized bed centrifuge

Abstract

Intensified high cell density (HCD) processes for the production of therapeutic proteins have already reached cell concentrations up to 100 million cells/mL (> 300 g/L wet cell weight, WCW). The clarification of such HCD processes using technologies established for conventional fed-batch processes (< 100 g/L WCW) is faced with low biomass loading capacities and relatively low product recoveries, which limit their applicability. This and the growing demand for single-use (SU) technologies increases the pressure to develop scalable, robust and cost-effective SU based HCD clarification solutions.Therefore, in this study a SU fluidized bed centrifuge (FBC) system was investigated to clarify HCD broths with 100 million cells/mL from a monoclonal antibody production process. FBC parameters including process volumes and flow rates during the FBC operating steps were optimized so that an almost complete removal of biomass and product recoveries of 95 % were achieved. In addition, the mild centrifugation conditions were demonstrated by the superior cell viability for the separated cells while minimizing release of host cell impurities. Furthermore, a post-centrifugal filter screening showed acceptable turbidities below 5 NTU of the sterile filtrate. This concept of FBC with subsequent filtration offers excellent potential to achieve a robust HCD clarification process solution.