(499g) Downstream Processing in Biomanufacturing: Multimodal Chromatography, Affinity Precipitation and Integrated Bioprocessing

This talk will present recent developments in our lab in three areas of downstream bioprocessing; multimodal chromatography, affinity precipitation and integrated bioprocessing. Targeted experiments with different domains of mAbs with varying properties, biophysics, thermodynamics and molecular simulations will be used to shed light on the importance of protein surface cluster properties and multimodal ligand hydration and flexibility on creating selective separations. Hierarchical clustering is employed to examine preferred ligand conformations and the impact of water and dewetting phenomena in multimodal systems. NMR and molecular simulations will be used to examine the molecular basis for multimodal ligand binding to the FC region of mAbs both in solution and immobilized on SAM surfaces. In addition, pH and ligand chemistry will be employed to shift domain dominance in mAb multimodal chromatography from (Fab)2 interactions to Fc binding. Affinity precipitation using smart biopolymers for the simultaneous recovery and purification of both mAb and non-mAb biologics will then be presented. The use of ELP-Z for mAb purification as well as the development of efficient processes based on this approach will be presented with industrial feed stocks. The extension of this approach using ELP-affinity peptides as well as the application to a new class of biologics will also be discussed. Finally, results will be given on a novel approach for the rapid development of integrated downstream biomanufacturing processes for biological products from Pichia Pastoris. This approach employs linear gradient screens in concert with high-throughput uplc analytics and custom software to identify sequences of orthogonal chromatography steps likely to recover and purify the product while also being amenable to integrated processing and a continuous flowpath. To illustrate the power of this novel approach, the rapid development of 3-step purification processes for three commercial biological products will be shown.