Antibody purification using porous metal-chelated monolithic columns

A novel monolithic material was developed to obtain efficient and cost-effective purification of IgG from human plasma. The porous monolith was obtained by bulk polymerization in a glass tube of 2-hydroxyethyl methacrylate (HEMA) and N-methacryloyl-(L)-histidine methyl ester (MAH). The poly(HEMA-MAH) monolith had a specific surface area of 214.6 m(2)/g and was characterized by swelling studies, porosity measurement, FTIR, scanning electron microscopy, and elemental analysis. Then the monolith was loaded with Cu2+ ions to form the metal chelate. Poly(HEMA-MAH) monolith with a swelling ratio of 74% and containing 20.9 mu mol MAH/g was used in the adsorption/desorption of IgG from aqueous solutions and human plasma. The maximum adsorption of IgG from an aqueous solution in phosphate buffer was 10.8 mg/g at pH 7.0. Higher adsorption was obtained from human plasma (up to 104.2 mg/g), with a purity of 94.1%. It was observed that IgG could be repeatedly adsorbed and desorbed with the poly(HEMA-MAH) monolith without significant loss of adsorption capacity. (c) 2006 Wiley Periodicals, Inc.