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. A novel approach has been developed to probe kinetics of protein-MM ligand interactions using quartz crystal microbalance with dissipation (QCM-D). QCM-D was used to obtain real time mass adsorption and desorption data of proteins binding to self-assembled monolayers (SAMs) of MM ligands that mimic a chromatographic resin surface, in a continuous flow mode. A systematic approach has been employed to investigate effects of ligand density by creating SAMs of different MM ligand densities on gold coated quartz crystals. In the present study, two different MM cation exchange ligand systems, namely the �Nuvia cPrime� like ligand and the �Capto MMC� like ligand were employed to delve into the differences in protein binding kinetics. A quantitative understanding of protein adsorption and desorption kinetics is obtained under the different conditions employed in this study. Moreover, binding behavior of proteins under different fluid phase conditions is examined in order to delineate the fundamental reasons for protein selectivity. The insights gained from these studies could potentially reveal the importance of ligand density and mobile phase modifiers for protein selectivity in multimodal systems which could have a significant impact on the development of more selective bioprocesses.

Investigation of protein selectivity in multimodal chromatography using quartz crystal microbalance with dissipation

247th National Meeting (2014)

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