EFFECTS OF COUPLING METHOD AND LIGAND CONCENTRATION ON ADSORPTION EQUILIBRIUM IN IMMUNO-AFFINITY CHROMATOGRAPHY

Abstract

The effects of ligand concentration and coupling method on the adsorption equilibrium in immuno-affinity chromatography were studied by use of polyclonal anti-BSA antibody and F_ab fragments. Immobilized IgG and F_ab fragments showed similar adsorption behavior, and the adsorption equilibrium was not affected by the coupling method or the concentration of immobilized ligands. However, the binding ratio, defined as the ratio of mole antigen adsorbed to mole binding site, depended on the coupling method and decreased with increasing ligand concentration.