KINETIC-STUDY OF THE ADSORPTION OF HUMAN SERUM-ALBUMIN ON IMMOBILIZEDANTIBODY USING THE SPLIT-PEAK EFFECT IN IMMUNOCHROMATOGRAPHY

The split-peak effect was used to determine the association rate const ant of the antigen-immobilized antibody reaction. The amount of immobi lized human serum albumin antibody on the chromatographic support was varied in order to find the optimal conditions to reduce the mass tran sfer contribution in the stagnant mobile phase fluid and measure the e ffective association rate constant of human serum albumin with the imm obilized antibody. Kinetic studies as a function of how-rate demonstra te the validity of the method consisting in determining the associatio n rate constant from measurements performed on columns of various capa cities. These experiments show that limitations due to mass transfer t o the surface of the adsorbent are minimized at high flow-rates and fo r a low density of immobilized ligand.