Biointeraction analysis of immobilized antibodies and related agents by high-performance immunoaffinity chromatography

A method is described based on high-performance immunoaffinity chromatography for examining the interactions of immobilized antibodies or related binding agents with their targets. It is shown how this method can be used to obtain information on the binding, elution and regeneration kinetics of immobilized binding agents, such as those used with immunoaffinity supports. The theory behind this approach is briefly described and it is demonstrated how both the kinetic and thermodynamic properties of a biointeraction can be determined experimentally through this method. Several applications are used to illustrate this technique, including antibody–antigen interactions and the binding of aptamers with their targets in the presence of silica-based supports. The same approach can be adapted for use with other types of targets, binding agents and support materials.

► A chromatographic method is described for biointeraction studies on solid supports.
► Kinetic and thermodynamic information can be obtained through this method.
► This method can study immobilized antibodies, aptamers and other binding agents.
► This method can provide data on target adsorption or elution and ligand regeneration.