Magnetic hydrophobic affinity nanobeads for lysozyme separation

AKTAŞ UYGUN D., Karagozler A. A., Akgol S., DENİZLİ A.

MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS, vol.29, no.7, pp.2165-2173, 2009 (SCI-Expanded) identifier identifier


N-Methacryloyl-l-phenylalanine (MAPA) containing poly(2-hydroxyethylmethacrylate) based magnetic [mag-poly(HEMA-MAPA)] nanobeads was prepared for lysozyme purification form chicken egg white. MAPA was synthesized by reacting methacryloyl chloride with l-phenylalanine methyl ester and provided hydrophobic functionality to the nanobeads. Size of mag-poly(HEMA-MAPA) nanobeads was 386 nm and obtained by surfactant free emulsion polymerization of HEMA and MAPA having a specific surface area of 580 m(2)/g. Mag-poly(HEMA-MAPA) nanobeads were characterized by FTIR, AFM, TEM, ESR, and elemental analysis. Lysozyme adsorption experiments were investigated under different conditions in batch system (i.e., medium pH, protein concentration, temperature, salt type). Lysozyme adsorption capacity of magpoly(HEMA) and mag-poly(HEMA-MAPA) nanobeads from aqueous solutions was estimated as 24 and 517 mg/g, respectively. Lysozyme molecules were desorbed with 50% ethylene glycol solution with 98% recovery. It was observed that mag-poly(HEMA-MAPA) nanobeads can be used without significant decrease in lysozyme adsorption capacity after ten adsorption-desorption cycles. Mag-poly(HEMA-MAPA) nanobeads was used for the purification of lysozyme from chicken egg white. Purity of lysozyme was estimated by SDS-PAGE. (C) 2009 Elsevier B.V. All rights reserved.