Co‐operative binding interactions in affinity chromatography: Theoretical considerations

Abstract

A theoretical relationship has been developed to allow the effect of free ligand concentration on the capacity of an affinity Chromatography matrix to be determined where the protein adsorbed shows co-operative binding. Computer simulations using literature values for association constants show that under optimal conditions resin capacity can be increased significantly in the presence of a small but finite concentration of free ligand. The model also allows prediction of the soluble ligand concentration required for biospecific elution. The results obtained suggest the possibility of a new elution technique, “reverse biospecific elution,” that reduces the amount of free ligand required to effect elution.