Computer Simulations of Gel Filtration Chromatograms of Rapidly Self-Associating Systems

A computer simulation method for gel filtration chromatography (GFC), based on plate theory and mass action law, is developed for rapidly self-associating systems and compared with asymptotic theory. For ovalbumin, a non-associable protein, whose elution volume increases linearly with increasing concentration, the leading boundary is predicted to be broader than the trailing. Computer simulations are used to estimate the aggregation numbers and the equilibrium constants for dimerization of carbonylhemoglobin and hexamerization of α-chymotrypsin. The concentration, Cmin, at the plateau region of the trailing boundary is shown to be close to cmc by asymptotic theory. The present computer simulation and derivative GFC patterns serve to estimate monomer concentration and micellar size of surfactants from analysis of GFC data.