Abstract: A high performance affinity chromatography medium used for separation of antithrombin Ⅲ was prepared. Monodisperse poly(glycidyl methacrylate) beads(PGMA) were employed as the affinity support. Heparin was linked covalently to amino-PGMA-beads, which was prepared by amination of PG-MA. The resultant affinity media present a minimal non specific interaction with proteins and the protein recovery was 96.9% and 94.4% for antithrombin Ⅲ and human serum albumin, respectively. Its affinity capacity for antithrombin Ⅲ was 12 mg/g beads. The apparent dissociation constant between antithrombin Ⅲ and immobilized heparin was 1.96×10^-5 mol/Lwhich was evaluated by front chromatography. The affinity medium was used to separate antithrombin Ⅲ from human plasma.

Key words: High performance affinity chromatography, Heparin, Antithrombin Ⅲ