Operating Protein A chromatography in continuous multi-column mode significantly improves resin capacity utilization and productivity compared with that in batch mode. However, the Protein A resin lifetime performance in continuous multi-column mode is still not well understood. As two or more columns always stay connected in the load zone in continuous mode, it is possible that the resin lifetime behaves differently than that of a single column in batch mode. We performed a resin lifetime study with a novel Protein A resin, running in a continuous 3-column mode, for 150 cycles. The resin binding capacity and yield results over cycles were compared with those from a single column running in batch mode for 150 cycles. Chromatography general rate model coupled with Langmuir adsorption isotherm was used to fit the breakthrough curves to investigate changes in resin properties, including maximum adsorption capacity and protein diffusivity coefficients, to understand the underlying mechanisms of resin fouling and ligand degradation. This knowledge facilitates predicting product loss of continuous multi-column Protein A chromatography after a certain number of cycles, and in developing corresponding strategies such as adjustable product loading and effective resin cleaning methods.

Mechanistic understanding of protein A resin lifetime in continuous multi-column mode

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.