(640a) Fundamental Studies in Multimodal Chromatography Using Protein Libraries, NMR and Multi-Scale Simulations

The design of novel multimodal chromatographic requires a fundamental understanding of the binding of the ligands involved in these processes to protein surfaces. In this presentation a variety of experimental and theoretical approaches are employed to provide insight into the binding mechanisms involved in these processes and to exploit them for the design of chromatographic systems with unique selectivities. Studies were carried out with various combinations of protein libraries, stationary phase ligands and fluid phase compositions. Protein libraries included protein ladders as well as homologous libraries of protein variants and commercial protein libraries. Mixed mode ligands included a range of chemically diverse ligands and were selected from libraries designed for selective binding to specific protein classes. The tools employed in this work include high throughput batch and column chromatography, NMR and simulations. Nuclear Magnetic Resonance was conducted on a range of protein/stationary phase ligand mixtures to verify the binding to targeted proteins and the location of the binding event on both the ligand and protein surfaces. Molecular dynamic simulations were conducted to corroborate the NMR results and to provide further insight into the binding and preferred protein-binding orientations on the chromatographic surfaces. Finally, this data is integrated into multi-scale models to aid in the development of predictive models of multimodal preparative chromatography for the purification of protein therapeutics.