ABSTRACT
The measurement of the equilibrium-binding constant between an immobilized-affinity ligand and a solute is relatively straightforward because the biospecific interaction of interest is almost always the major cause of retention in the affinity-chromatographic column. The purpose of the theory and computer simulation sections is to help define the limitations of each of the methods. This chapter discusses the theory of rate-constant measurement under isocratic-elution conditions. The isocratic method and two other methods are discussed from a more practical point of view using experimental data, example calculations, and computer simulations. The purpose of the theory and computer simulation sections is to help define the limitations of each of the methods. These limitations are then used in the example calculations to determine the necessary experimental conditions and whether the desired measurement can even be made. Computer-generated peak profiles are useful for predicting peak shapes and other parameters which cannot be calculated directly from theory.
