Antibody purification from human plasma by metal‐chelated affinity membranes

Abstract

AbstractThe aim of this study is to investigate in detail the feasibility of poly(2‐hydroxyethyl methacrylate‐N‐methacryloyl‐(L)‐histidine methyl ester), PHEMAH membranes for purification of immunoglobulin G (IgG) from human plasma. PHEMAH membranes were prepared by photo‐polymerization technique. Then, Zn2+, Ni2+, Co2+, and Cu2+ ions were chelated directly on the PHEMAH membranes. Elemental analysis assay was performed to determine the nitrogen content and polymerized MAH was calculated as 168.5 μmol/g. The nonspecific IgG adsorption onto the plain PHEMA membranes was negligible (about 0.25 mg/mL). A remarkable increase in the IgG adsorption capacities were achieved from human plasma with PHEMAH membranes (up to 68.4 mg/mL). Further increase was observed with the metal‐chelated PHEMAH membranes (up to 118 mg/mL). The metal‐chelate affinity membranes allowed the one‐step separation of IgG from human plasma. The binding range of metal ions for surface histidines from human plasma followed the order: Cu2+ > Ni2+ > Zn2+ > Co2+. Adsorbed IgG was eluted using 250 mM EDTA with a purity of 94.1%. IgG molecules could be repeatedly adsorbed and eluted with the metal‐chelated PHEMAH membranes without noticeable loss in their IgG adsorption capacity. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012