Recent studies in our group have shown that the combination of displacers and mobile phase modifiers in multimodal (MM) systems can create unique selectivity windows for protein separations. There is a need to understand the fundamental reasons that govern such selective behavior. Quartz crystal microbalance with dissipation (QCM- D) was used to obtain real time mass adsorption and desorption data of proteins binding to self-assembled monolayers (SAM) of MM ligands that mimic a chromatographic resin surface, in a continuous flow mode. A systematic approach has been employed to extract kinetic rates of protein desorption by investigating the binding behavior of proteins under different fluid phase conditions in order to delineate the fundamental reasons for selectivity. The phenomena of changeover in binding behavior from selective desorption to selective displacement has been studied. The insights gained from these studies reveal the nature of selectivity created by mobile phase modifiers and displacers in MM systems.

Probing protein selectivity in multimodal chromatography using quartz crystal microbalance

245th National Meeting (2013)

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