Impact of Higher Order Structure on Ligand Binding

This technical report investigates the relationship between higher order structure changes and analyte ligand binding, utilizing Biolayer Interferometry (BLI) and Microfluidic Modulation Spectroscopy (MMS). The study focuses on how variations in protein concentration and functionality affect binding interactions, particularly in the context of formulation development. It outlines the methodologies employed, including the evaluation of binding activity and hydrophobic surface binding for aggregated molecules. The report emphasizes the importance of accurate buffer matching to ensure reliable results, as minor variations in buffer concentrations can lead to unreliable data. Additionally, the report discusses the structural stability of proteins in relation to their binding activity, detailing how different excipients and conditions can influence protein conformation and function. The findings underscore the potential of combining BLI and MMS technologies for a deeper understanding of protein behavior in various formulations, enhancing the assessment of biologics stability and functionality.