Poorly packed columns can have serious consequences on the performance of chromatography and efforts to understand the impact of different methods of compression on protein separation is limited in the literature. This thesis examines the effects of hydrodynamic and mechanical compression on process chromatography. This results in higher productivity and could contribute to cost reduction. High throughput resins can be used for purification of high volume and high concentration feedstock in limited time. Ligand accessibility was therefore largely enhanced, allowing proteins to interact and bind under harsh conditions. Moreover, they maintained mainly high capacities even with extremely high flow velocities. Structure-extended resins showed higher binding capacities compared to resins with conventional ligands directly attached to the matrix. By systematic frontal analysis and selectivity determination at different flow rates, we suggested an independent evaluation of major anion exchangers to facilitate media selection, and investigated the relationship between (i) surface modification and (ii) chromatographic performances. New ion exchangers have tailored matrices providing low backpressure and allowing high flow velocity. Chromatographic performances are highly influenced by operational parameters.
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