Chromatographic study of the thermodynamic and kinetic characteristics of silica-bound concanavalin A

A method has been developed for determining the rate constants for dissociation of sugar derivatives from silica-immobilized concanavalin A using high-performance liquid affinity chromatography. The rate constants observed are ca. ten-fold smaller than those reported in solution. Using 10-μm silica particles, it has been shown that slow dissociation of the sample from the immobilized affinity ligand is the predominant factor responsible for the larger plate heights observed in this system. The effect of several chromatographic variable (temperature, mobile phase modifiers and particle size) on the observed dissociation rate constant and peak centroid have been evaluated.