Panmap

Abstract

Multivalent binding is a major feature of many biological systems including the interactions of antibodies and antigens, the understanding of which requires accounting for how multiple binding states make up the overall affinity and avidity to determine the strength and specificity of an interaction. Traditional affinity measurements, such as ELISA, provide a limited view of binding interactions but not the means to account for multivalence. We therefore developed the Patterned Antigen Nanostructure-derived Multivalent Avidity Profiling (PANMAP) method, which enables affinity measurement combined with a concise representation that considers multivalence. We discovered that distance between ligands determines the equilibrium binding of antibodies, and that it is defined by a near-distance competitive exclusion regime, a far-distance non-bivalent regime, and an intermediate optimal bivalent regime - each with different outcomes, highlighting the importance of ligand patterns in biological systems where multivalent binding is possible. This finding shows how this simple tool to measure multivalent equilibrium binding could play a role in quickly determining a detailed picture of binding for a variety of systems, enabling better insight into biological processes and greater control during commercial engineering of binding partners.