Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels

BERELİ N., ANDAÇ A. M., Baydemir G., Say R., Galaev I. Y., DENİZLİ A.

JOURNAL OF CHROMATOGRAPHY A, vol.1190, no.1-2, pp.18-26, 2008 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 1190 Issue: 1-2
  • Publication Date: 2008
  • Doi Number: 10.1016/j.chroma.2008.02.110
  • Journal Name: JOURNAL OF CHROMATOGRAPHY A
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.18-26
  • Keywords: cryogels, molecular imprinting, molecular recognition, lysozyme purification, protein adsorption, affinity binding, SOLID-PHASE EXTRACTION, IMMUNOGLOBULIN PRODUCTION, METHACRYLATE) BEADS, STATIONARY-PHASE, CADMIUM REMOVAL, AFFINITY, LYSOZYME, SEPARATION, PRECONCENTRATION, POLYMERS
  • Anadolu University Affiliated: Yes

Abstract

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)-histiclinemethylester (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 1 M 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. (C) 2008 Elsevier B.V. All rights reserved.