Development of mL-scale pseudo-perfusion methodologies for high-throughput early phase development studies

Small-scale devices enable rapid development and optimization, while many exist for fed-batch cultures, there are few comparable high cell density perfusion cultures. To address this gap, pseudo-perfusion methodologies were developed in microwell plates at mL-scale capable of achieving the specific characteristics culture, including density, retention increased productivity. Pseudo-perfusion was achieved via sedimentation or centrifugation 24 prior to media exchange 1–2 times daily, generating separation efficiencies higher than 90%. Media exchanges commenced on day 3 rates 0.5–1.8 vessel volumes per (VVD). resulted maximum viable densities (VCDs) up 42 × 106 cells mL-1, 4.2-fold greater Volumetric productivities by 1.9-fold, industrially relevant productivities. Similar performance observed between methodologies, with minor deviations attributed elongated manipulation lower packed when used. The plate (MWP) experiments validated 5 L scale showed comparability metabolite growth profiles. work presents a robust tool high-throughput studies which is sensitive changes composition rate, could be reliably used initial screening