Preparation of a novel metal‐chelate affinity beads for albumin isolation from human plasma

In this study, we developed a novel approach to obtain a high protein‐adsorption capacity utilizing 2‐methacryloylamidohistidine (MAH) as a biollgand. MAH was synthesized by reacting methacryloyl chloride and histidine. Spherical beads, with an average size of 150–200 μm, were obtained by the radical suspension polymerization of MAH, ethyleneglycol dimethacrylate (EGDMA), and 2‐hydroxyethyl methacrylate (HEMA) conducted in an aqueous dispersion medium. p(EGDMA–HEMA–MAH) beads had a specific surface area of 17.6 m²/g. The synthesized MAH monomer was characterized by NMR. p(EGDMA–HEMA–MAH) beads were characterized by a swelling test, FTIR, and elemental analysis. Then, Cu(II) ions were incorporated into the beads and Cu(II) loading was found to be 0.96 mmol/g. These beads, with a swelling ratio of 65% and containing 1.6 mmol MAH/g, were used in the adsorption/desorption of human serum albumin (HSA) from both aqueous solutions and human serum. The adsorption of HSA onto p(EGDMA–HEMA–MAH) was low (8.8 mg/g). Cu(II) chelation onto the beads significantly increased the HSA adsorption (56.3 mg/g). The maximum HSA adsorption was observed at pH 8.0 Higher HSA adsorption was observed from human serum (94.6 mg HSA/g). Adsorptions of other serum proteins were obtained as 3.7 mg/g for fibrinogen and 8.5 mg/g for γ‐globulin. The total protein adsorption was determined as 107.1 mg/g. Desorption of HSA was obtained using a 0.1M Tris/HCI buffer containing 0.5M NaSCN. High desorption ratios (to 98% of the adsorbed HSA) were observed. It was possible to reuse Cu(II)‐chelated p(EGDMA–HEMA–MAH) beads without significant decreases in the adsorption capacities. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 2840–2847, 2003