CHROMATOGRAPHIC STUDY OF THE ADSORPTION-KINETICS OF ALBUMIN ON MONOCLONAL AND POLYCLONAL IMMUNOADSORBENTS

High-performance liquid chromatography (HPLC) has been used to study t he adsorption kinetics of proteins on immunoadsorbents. The adsorption rate constant of human serum albumin (HSA) on monoclonal and polyclon al anti-HSA antibodies immobilized on a silica HPLC support was determ ined by saturating the column with repeated pulse injections. Studies on polyclonal immunoadsorbents of different capacities enable evaluati on of the contribution of transport to the binding sites. The adsorpti on properties of two different monoclonal anti-HSA antibodies immobili zed on a chromatographic support were characterized by different appro aches. The location of the epitope on the HSA molecule was determined by enzyme-linked immunoassay (ELISA) with albumin fragments. The chrom atographic method was used to determine the column capacity and the ad sorption rate constant of HSA on the immunoadsorbent. To compare the a ffinity of the antibodies for the antigen, an indirect ELISA method wa s used to determine the equilibrium constant of antigen-antibody assoc iation in solution.