Development of a high-throughput, automated mammalian perfusion cell culture process development tool

Continuous bioprocessing of therapeutic proteins is becoming an increasingly prevalent process mode in research and development as well as manufacturing organizations.  Perfusion processes offer high volumetric productivities, short therapeutic protein residence time, and long term operation.  As a downside, perfusion processes require specialized capital equipment (bioreactor and cell separation device), large media volumes and significant personnel resources.  These requirements lead to lengthy and costly development timelines.  This presentation describes the development of automated high-throughput workflow aimed at improving the efficiency of perfusion cell culture process development.  This developmental tool replicates high-density cell cultivation by maintaining cultures in 50 ml TPP bioreactor tubes and employing automated liquid handling via a custom Tecan Freedom Evo liquid handler.  A daily manual centrifugation step is used to mimic the cell retention and media exchange of a typical bioreactor perfusion process.  This developmental tool can be used for cell line selection and media development applications.  Experiments can be completed in under seven days and utilize less than 50 ml of media per condition, which represents over a 4-fold reduction in culture time and 1000-fold reduction in media usage for a typical perfusion processes. Daily samples can be drawn into 96 well plates for direct integration with high-throughput analytical assays, generating rapid results for feedback control or identification of influential observations.  Data trends observed in the small scale development tool can be correlated with verification runs in benchtop perfusion bioreactors.