Analytical affinity chromatography: II. Rate theory and the measurement of biological binding kinetics

Abstract

Affinity chromatography can be used to measure equilibrium constants and kinetics of biological interactions. The local-equilibrium theory presented in the preceding paper is extended to include mass transfer and kinetic effects. Solutions for both zonal and frontal elution are presented. For highly nonlinear isotherms, the frontal elution method is preferred. Experiments with bovine serum albumin binding to immobilized Reactive Blue show that the binding kinetics inside the porous gel are several orders of magnitude slower than typical biological binding reactions in solution. The temperature dependence of the kinetic constants indicate that the binding may still be diffusion-controlled.