title: Polyethyleneimine assisted-two-step polymerization to develop surface imprinted cryogels for lysozyme purification.
abstract: Surface imprinting strategy is one of the promising approaches to synthesize plastic antibodies while overcoming the problems in the protein imprinting research. In this study, we focused our attentions on developing two-step polymerization to imprint on the bare surface employing polyethyleneimine (PEI) assisted-coordination of template molecules, lysozyme. For this aim, we firstly synthesized poly(2-hydroxyethyl methacrylate-glycidyl methacrylate), poly(HEMA-GMA) cryogels as a bare structure. Then, we immobilized PEI onto the cryogels through the addition reaction between GMA and PEI molecules. After that, we determined the amount of free amine (NH2) groups of PEI molecules, subsequently immobilized methacrylate functionalities onto the half of them and another half was used to chelate Cu(II) ions as a mediator between template, lysozyme and PEI groups. After the characterization of the materials developed by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and the micro-computed tomography (μCT), we optimized the lysozyme adsorption conditions from aqueous solution. Before performing lysozyme purification from chicken egg white, we evaluated the effects of pH, interaction time, the initial lysozyme concentration, temperature and ionic strength on the lysozyme adsorption. Moreover, the selectivity of surface imprinted cryogels was examined against cytochrome c and bovine serum albumin (BSA) as the competitors. Finally, the mathematical modeling, which was applied to describe the adsorption process, showed that the experimental data is very well-fitted to the Langmuir adsorption isotherm.
authors.affiliation: Hacettepe University, Department of Chemistry, Ankara, Turkey; Hitit University, Osmancık Ömer Derindere Vocational Higher School, Department of Property Protection and Safety, Çorum, Turkey.
authors.forename: Kadir
authors.initials: K
authors.lastname: Erol
authors.affiliation: Hacettepe University, Department of Chemistry, Ankara, Turkey; Hitit University, Scientific Technical Research and Application Center, Çorum, Turkey.
authors.forename: Kazım
authors.initials: K
authors.lastname: Köse
authors.affiliation: Hacettepe University, Department of Chemistry, Ankara, Turkey. Electronic address: lokman@hacettepe.edu.tr.
authors.forename: Lokman
authors.initials: L
authors.lastname: Uzun
authors.affiliation: Eskişehir University, Department of Chemistry, Eskişehir, Turkey.
authors.forename: Rıdvan
authors.initials: R
authors.lastname: Say
authors.affiliation: Hacettepe University, Department of Chemistry, Ankara, Turkey.
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D059985:Cryogels / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D009113:Muramidase / Q000302:isolation & purification*; D011094:Polyethyleneimine / Q000737:chemistry*; D058105:Polymerization; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D059985:Cryogels; D011094:Polyethyleneimine; D009113:Muramidase
keywords: Cryogel; Lysozyme; Polyethyleneimine; Surface imprinting; Two-step polymerization