title: Antibody-imprinted membrane adsorber via two-step surface grafting.
abstract: In this work, a recently established, novel two-step imprinting strategy combining surface imprinting and scaffold imprinting was applied successfully to prepare a molecularly imprinted polymer (MIP) adsorber for immunoglobulin G (IgG). Track-etched polyethylene terephthalate (PET) membranes with previously introduced aliphatic C-Br groups as initiator on the pore surface were used to prepare first a functional polymer scaffold, grafted poly(methacrylic acid), via surface-initiated atom transfer radical polymerization (SI-ATRP). After template protein (IgG) binding to the scaffold, UV-initiated cross-linking copolymerization of acrylamide and methylenebisacrylamide (MBAA) as second step lead to a grafted MIP hydrogel layer. The influences of the three independent parameters, scaffold chain length by SI-ATRP time, degree of cross-linking of the MIP layer by MBAA content, and grafted MIP layer thickness by UV irradiation time, were studied to optimize protein binding capacity and selectivity. The results were also compared to previously obtained data for lysozyme imprinting using the same method, and significant effects of protein size on imprinting efficiency could be identified. The best IgG MIP membrane adsorber was then used to separate IgG from mixtures with human serum albumin (HSA), demonstrating IgG binding capacities and eluted IgG purities, which were almost independent of the excess of HSA. The results of this study are a significant extension of the scope of molecular imprinting toward large target bionanoparticles. The transfer of the approach from the model PET to other base membranes with higher specific surface area is straightforward, and the resulting affinity materials would, in principle, be suited for "capturing" of an antibody from a complex mixture.
authors.affiliation: Lehrstuhl für Technische Chemie II, Universität Duisburg-Essen , 45117 Essen, Germany.
authors.forename: Dongxu
authors.initials: D
authors.lastname: Yin
authors.forename: Mathias
authors.initials: M
authors.lastname: Ulbricht
mesh_terms: D000178:Acrylamides / Q000737:chemistry; D000327:Adsorption; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D008567:Membranes, Artificial; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D011092:Polyethylene Glycols; D011093:Polyethylene Terephthalates; D011109:Polymethacrylic Acids / Q000737:chemistry; D011485:Protein Binding; D012709:Serum Albumin / Q000302:isolation & purification; D013499:Surface Properties
chemical_list: D000178:Acrylamides; D007074:Immunoglobulin G; D008567:Membranes, Artificial; D011093:Polyethylene Terephthalates; D011109:Polymethacrylic Acids; D012709:Serum Albumin; C030613:polymethacrylic acid; D011092:Polyethylene Glycols; D009113:Muramidase; C021221:N,N'-methylenebisacrylamide