Mobile Phase Modifier Effects In Multimodal Chromatography: Fundamental Understanding and Creation of Selectivity Windows

A multifaceted approach consisting of chromatography, nuclear magnetic resonance, and isothermal titration calorimetry was used to provide fundamental insight into protein adsorption in multimodal chromatographic systems.  A wide experimental space was examined using column and high-throughput batch chromatography experiments with a variety of mobile phase modifiers.  These experimental techniques were used in conjunction with molecular simulations to understand protein-ligand interactions at the molecular level. The results demonstrate how the appropriate use of mobile phase modifiers and multimodal resins can enable unique separations to be performed that cannot be achieved with traditional methods.