Multi-modal chromatographic systems offer improved selectivity over traditional separation methods such as ion exchange or hydrophobic interaction chromatography.   In order to further improve the selectivity in these systems, a variety of mobile phase modifiers were employed to probe the binding mechanisms of protein libraries to several multi-modal resins.  Experiments were carried out using high throughput batch experiments as well as column experiments with varying classes of mobile phase additives, pH, salt types and gradients.   The effect of mobile phase modifiers on multimodal ligand protein binding was also explored using NMR with isotopically labeled proteins and free floating forms of multi-modal ligands in the presence of these mobile phase modifiers.   The results clearly demonstrate how the appropriate use of mobile phase modifiers and multi-modal resins can enable unique separations to be performed that can not be achieved with traditional methods.

Improving the selectivity in multimodal chromatographic systems with mobile phase modifiers: Fundamental and applied studies

239th National Meeting (2010)

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