Determination of kinetic parameters of epitope-paratope interaction based on solid phase binding: An inexpensive alternate to biospecific interaction analysis.

ABSTRACT

A method has been developed for biospecific interaction analysis between antigen and antibody using solid phase binding approach. Real time kinetics between monoclonal antibody and human chorionic gonadotropin have been studied. Kinetic constants of the bimolecular reaction are determined. Affinity constants measured by several independent methods have been found to be relatively consistent. Convenient and simple procedures to determine affinity constant, Kon and Koff of monoclonal antibody-human chorionic gonadotropin interaction using binding of [125I]hCG to immobilized monoclonal antibody are presented. Values obtained compare well with those obtained using surface plasmon resonance technology,making this method a viable alternative.