This article reviews the progress of a personal endeavour to develop chromatography as a quantitative procedure for the determination of reaction stoichiometries and equilibrium constants governing protein interactions. As well as affording insight into an aspect of chromatography with which many protein chemists are unfamiliar, it shows the way in which minor adaptations of conventional chromatographic practices have rendered the technique one of the most powerful methods available for the characterization of interactions. That pathway towards quantification is followed from the introduction of frontal gel filtration for the study of protein self-association to the characterization of ligand binding by the biosensor variant of quantitative affinity chromatography.
- analytical exclusion chromatography
- ligand binding
- protein interactions
- quantitative affinity chromatography
- solute self-association
Intermolecular Associations in 2D and 3D, a Biochemical Society Focused Meeting held at University of Nottingham, 19–20 June 2003
- © 2003 Biochemical Society