(263c) Evaluation of Protein Adsorption in Multimodal Chromatographic Systems

Multimodal chromatographic systems offer improved selectivity over traditional separation methods such as ion exchange or hydrophobic interaction chromatography. However, the complex nature of the interactions involved in multimodal systems hinders the optimal use of these materials. High-throughput batch chromatographic experiments were employed to evaluate the performance and selectivity of a library of multimodal materials. Column experiments with varying classes of mobile phase additives, pH, and salt types provided further insight into the protein binding mechanisms with several resins. Multimodal ligand-protein binding was also explored using nuclear magnetic resonance experiments with isotopically-labeled proteins and multimodal ligands in the presence of these mobile phase modifiers. The combination of column, batch, and free solution experiments greatly enhanced the fundamental understanding of protein adsorption in multimodal chromatographic systems. These results have important implications for methods development applications and future multimodal ligand design.