Coiled Flow Inverter Reactor (CFIR) has recently been explored for facilitating continuous operation of several unit operations involved in downstream processing of biopharmaceuticals such as viral inactivation and protein refolding. In this study, the application of CFIR for continuous precipitation of clarified cell culture supernatant has been explored. The pH based precipitation was optimized in the batch mode and then in the continuous mode in CFIR using a design of experiments (DOE) study. Improved clearance of host cell DNA (52X vs. 39X in batch), improved clearance of host cell proteins (HCP) (7X vs. 6X in batch) and comparable recovery (90 % vs. 91.5 % in batch) were observed along with 6 times higher productivity. To further demonstrate wider applicability of CFIR in performing continuous precipitation, two more case studies involving use of two different precipitation protocols (CaCl<sub>2</sub> based and caprylic acid based) were also performed. In both cases, clearance of host cell DNA, HCP, and product recovery were found to be comparable or better in CFIR than in batch operations. Moreover, increase in productivity of 16 times (CaCl<sub>2</sub> based) and 8 times (caprylic acid based) was obtained for the two precipitation protocols respectively. The data presented in this study clearly demonstrates that CFIR can be seamlessly integrated into a continuous bioprocess train for performing continuous precipitation of clarified cell culture supernatant. To our knowledge this is the first report of such use.

Continuous precipitation of process related impurities from clarified cell culture supernatant using a novel Coiled Flow Inversion Reactor (CFIR)

253rd National Meeting (2017)

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