Physical Sciences and Mathematics Commons^™

Biointeraction Analysis By High-Performance Affinity Chromatography: Kinetic Studies Of Immobilized Antibodies, Mary Anne Nelson, Annette C. Moser, David S. Hage

David Hage Publications

A system based on high-performance affinity chromatography was developed for characterizing the binding, elution and regeneration kinetics of immobilized antibodies and immunoaffinity supports. This information was provided by using a combination of frontal analysis, split-peak analysis and peak decay analysis to determine the rate constants for antibody–antigen interactions under typical sample application and elution conditions. This technique was tested using immunoaffinity supports that contained monoclonal antibodies for 2,4-dichlorophenoxyacetic acid (2,4-D). Association equilibrium constants measured by frontal analysis for 2,4-D and related compounds with the immobilized antibodies were 1.7–12 × 10⁶ M⁻¹ at pH 7.0 and 25 °C. Splitpeak …