Conformational Changes during Adsorption and Chromatographic Separation of Bivalent Bi-Specific Antibodies - Biomolecular Perspectives

Proteins can exhibit conformational changes while adsorbed to the surface of resins used for chromatographic separations resulting in complex elution behaviors both for ion exchange chromatography and for hydrophobic interaction chromatography. Both unfolding with loss of secondary structure and reversible conformational changes have been reported. In either case, recovery and separation efficiency are impacted. Additionally, these phenomena complicate the development of mechanistic models since the classical, thermodynamics-based approaches for the description of protein surface interactions do not take such phenomena into account. In this talk, we present recent advances that we have made to gain a biomolecular understanding of the underlying phenomena and to model chromatographic elution of bivalent bi-specific antibodies whose multi-domain characteristics result in particularly complex elution behaviors.