Surface plasmon resonance is an effective technique to monitor the selectivity and affinity of biomolecular interactions. SPR can be used to analyze the association and dissociation rate coefficients and model biomolecular interactions kinetics. It also allows for equilibrium binding analysis, ligand specificity studies and analysis of dissociation rate constants.
Anti-SPR antibody has been widely used and improved the precision of classifying protein-protein interactions as well as studying small-molecule receptor ligand binding. However, lipid-protein interactions are still poorly served. The field of lipids is the future frontier in cell research.
SPR is an extremely advantageous technique for cell biologists. Newly identified proteins can be quickly and robustly screened for membrane affinity and lipid specificity. This technical perspective describes the conditions that are necessary to be successful with lipid protein interactions. It also highlights the unique lipid/protein interaction mechanisms that SPR has revealed.
This technical perspective is designed to give the reader a framework for confident and quantitative conclusions from SPR analysis on lipid-protein interactions. Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin and anti-fluorescein monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1.
The conjugation process caused only minor changes in the equilibrium dissociation constants for the antibody conjugates and their ligands. Comparing the initial binding rates of conjugates and their ligands to those of unmodified antibodies at different concentrations revealed that only a small percentage of antibody conjugates had been inactivated.