Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels.

Molecular imprinting is a method for making selective binding sites in synthetic polymers using a molecular template. The aim of this study is to prepare lysozyme-imprinted supermacroporous cryogels which can be used for the purification of lysozyme (Lyz) from egg white. N-Methacryloyl-(L)-histidinemethylester (MAH) was chosen as the metal-coordinating monomer. In the first step, Cu2+ was complexed with MAH and the lysozyme-imprinted poly(HEMA-MAH) [Lyz-MIP] cryogel were produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) in an ice bath. After that, the template (i.e., lysozyme) was removed using 0.05 M phosphate buffer containing 1M NaCl (pH 8.0). The maximum lysozyme adsorption capacity was 22.9 mg/g polymer. The relative selectivity coefficients of Lyz-MIP cryogel for lysozyme/bovine serum albumin and lysozyme/cytochrome c were 4.6 and 3.2 times greater than non-imprinted poly(HEMA-MAH) (NIP) cryogel, respectively. Purification of lysozyme from egg white was also monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 94% with recovery about 86%. The Lyz-MIP cryogel could be used many times without decreasing the adsorption capacity significantly.

[1]  A. Denizli,et al.  Dye-ligand affinity systems. , 2001, Journal of biochemical and biophysical methods.

[2]  A. Denizli,et al.  Binding of antibodies to concanavalin A-modified monolithic cryogel , 2006 .

[3]  Z. Cui,et al.  Protein purification by ultrafiltration with pre-treated membrane , 2000 .

[4]  R. Ansell.  MIP-ligand binding assays (pseudo-immunoassays) , 2001, Bioseparation.

[5]  G. Cheng,et al.  Synthesis of polyacrylamide gel beads with electrostatic functional groups for the molecular imprinting of bovine serum albumin , 2006, Analytical and bioanalytical chemistry.

[6]  A. Denizli,et al.  A Novel Affinity Support Material for the Separation of Immunoglobulin G from Human Plasma , 2002 .

[7]  G. Demirel,et al.  pH/temperature - sensitive imprinted ionic poly(N-tert-butylacrylamide-co-acrylamide/maleic acid) hydrogels for bovine serum albumin. , 2005, Macromolecular bioscience.

[8]  B. Mattiasson,et al.  Direct chromatographic capture of enzyme from crude homogenate using immobilized metal affinity chromatography on a continuous supermacroporous adsorbent. , 2003, Journal of chromatography. A.

[9]  A. Denizli,et al.  Iron removal from human plasma based on molecular recognition using imprinted beads , 2005 .

[10]  Filiz Yılmaz,et al.  Preconcentration of copper on ion-selective imprinted polymer microbeads , 2003 .

[11]  R. Huopalahti,et al.  Bioactive egg compounds , 2007 .

[12]  A. Denizli,et al.  Molecular recognition based cadmium removal from human plasma. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[13]  Adil Denizli,et al.  Molecularly imprinted ligand-exchange recognition assay of glucose by quartz crystal microbalance. , 2005, Biosensors & bioelectronics.

[14]  Yong Li,et al.  Protein recognition via surface molecularly imprinted polymer nanowires. , 2006, Analytical chemistry.

[15]  L. Ye,et al.  Polymers recognizing biomolecules based on a combination of molecular imprinting and proximity scintillation: a new sensor concept. , 2001, Journal of the American Chemical Society.

[16]  T. Rao,et al.  Tailored materials for preconcentration or separation of metals by ion-imprinted polymers for solid-phase extraction (IIP-SPE) , 2004 .

[17]  A. Denizli,et al.  Synthesis of cholesterol imprinted polymeric particles. , 2007, International journal of biological macromolecules.

[18]  Klaus Mosbach,et al.  Molecular imprinting: recent developments and the road ahead , 1999 .

[19]  K. Hidajat,et al.  Adsorption and desorption of lysozyme on nano-sized magnetic particles and its conformational changes. , 2004, Colloids and surfaces. B, Biointerfaces.

[20]  A. Denizli,et al.  Ion‐selective Imprinted Superporous Monolith for Cadmium Removal from Human Plasma , 2005 .

[21]  G. Cheng,et al.  Protein-imprinted soft-wet gel composite microspheres with magnetic susceptibility. II. Characteristics , 2006 .

[22]  Bo Mattiasson,et al.  Polymeric cryogels as promising materials of biotechnological interest. , 2003, Trends in biotechnology.

[23]  K. Mosbach,et al.  Synthesis and catalysis by molecularly imprinted materials. , 1999, Current opinion in chemical biology.

[24]  Adil Denizli,et al.  Ni(II) ion-imprinted solid-phase extraction and preconcentration in aqueous solutions by packed-bed columns , 2004 .

[25]  A. Denizli,et al.  Molecular imprinted particles for lysozyme purification , 2007 .

[26]  A. Denizli,et al.  L-histidine imprinted synthetic receptor for biochromatography applications. , 2006, Analytical chemistry.

[27]  A. Denizli,et al.  Monosize poly(glycidyl methacrylate) beads for dye-affinity purification of lysozyme. , 2006, International journal of biological macromolecules.

[28]  Yen Wah Tong,et al.  Defining the Interactions between Proteins and Surfactants for Nanoparticle Surface Imprinting through Miniemulsion Polymerization , 2008 .

[29]  A. Denizli,et al.  Poly(acrylamide-allyl glycidyl ether) cryogel as a novel stationary phase in dye-affinity chromatography , 2007 .

[30]  T. Sugahara,et al.  Lysozyme stimulates immunoglobulin production by human-human hybridoma and human peripheral blood lymphocytes , 1997, Cytotechnology.

[31]  Adil Denizli,et al.  Selective Separation of Uranium Containing Glutamic Acid Molecular-Imprinted Polymeric Microbeads , 2003 .

[32]  L. Ye,et al.  Use of molecularly imprinted polymers in a biotransformation process. , 1999, Biotechnology and bioengineering.

[33]  Cui,et al.  Lysozyme separation by hollow-fibre ultrafiltration. , 2000, Biochemical engineering journal.

[34]  A. Denizli,et al.  Poly(ethylene dimethacrylate-glycidyl methacrylate) Monolith as a Stationary Phase in Dye-Affinity Chromatography , 2004 .

[35]  Yan Lu,et al.  Coating lysozyme molecularly imprinted thin films on the surface of microspheres in aqueous solutions , 2007 .

[36]  B. Mattiasson,et al.  Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns. , 2002, Journal of chromatography. A.

[37]  Adil Denizli,et al.  Cibacron Blue F3GA incorporated magnetic poly(2‐hydroxyethyl methacrylate) beads for lysozyme adsorption , 2004 .

[38]  S. Striegler.  Designing selective sites in templated polymers utilizing coordinative bonds. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[39]  S. Banerjee,et al.  Experimental evaluation of preventive and therapeutic potentials of lysozyme. , 1992, Chemotherapy.

[40]  H. Bianco-Peled,et al.  Study of the interactions between protein-imprinted hydrogels and their templates. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[41]  B. Mattiasson,et al.  The potential of polymeric cryogels in bioseparation , 2001, Bioseparation.

[42]  T. Rao,et al.  Investigation of different polymerization methods on the analytical performance of palladium(II) ion imprinted polymer materials , 2005 .