Continuous biomanufacturing by means of perfusion processes enables constant product flow with short product residence times and constant product quality at high volumetric bioreactor productivities. Therefore, perfusion processes are promising tools for small, flexible, single-use biomanufacturing facilities of the future. Perfusion processes and media can be optimized for maximum viable cell densities, applied in N-1 perfusion operations for process intensification, or for minimal media consumption to reach highest product titers. Despite their potential, perfusion processes face challenges due to more complex process controls and lack of suitable small-scale models to develop high-performing perfusion media. To address the demands of a high-performing perfusion medium for a given cell line, we demonstrate a suitable screening workflow for optimal combination of commercially available HyClone basal media spiked with different HyClone Cell Boost supplements using a Design of Experiment approach. High-performing perfusion media compositions were derived from ActiPro or CDM4NS0 basal media and applied to perfusion processes in culture vessels at different scales starting from simulated TubeSpin pseudo-perfusion cultures, up to bioreactor perfusion runs in ReadyToProcess WAVE systems at 0.5 L or XDR-50 bioreactors at 40 L working volume, reaching cell densities above 200 × 106 c/mL and optimized cell-specific perfusion rates as low as 20 pL/c/d.

DoE-supported perfusion medium optimization for maximum cell densities and efficient media consumption

2019 Spring National Meeting

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