A framework based on the extended Wyman concept for analyzing the salt effects on the solute retention in high-performance affinity chromatography

摘要

The analysis of binding data of a ligand to a macromolecule in the presence of an additive can be classically formulated in terms of the linked functions of Wyman. In the case of a salt, this approach has been extended by Tanford such that the contributions of both salt and water are taken into account. In this paper, the extended Wyman theory was applied to high-performance affinity chromatography (HPAC) in order to define a general model describing the effects of the mobile-phase salts on the ligand binding. Various HPAC literature data, as well as our data concerning dansyl amino acid retention on a vancomycin stationary phase, were examined in relation to this model. From the results, this theoretical approach was considered to be adequate to describe accurately the salt dependence on solute retention. This work shows the importance of taking into account the effects of both ionic species and water in the investigation of relative contributions of the interactions involved in the ligand binding to immobilized receptor. [References: 36]