L-Histidine imprinted supermacroporous cryogels for protein recognition

BERELİ N., SAYLAN Y., UZUN L., Say R., DENİZLİ A.

SEPARATION AND PURIFICATION TECHNOLOGY, cilt.82, ss.28-35, 2011 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 82
  • Basım Tarihi: 2011
  • Doi Numarası: 10.1016/j.seppur.2011.08.011
  • Dergi Adı: SEPARATION AND PURIFICATION TECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.28-35
  • Anahtar Kelimeler: Cryogels, Molecular imprinting, Molecular recognition, Lysozyme purification, Affinity binding, SOLID-PHASE EXTRACTION, IMMUNOGLOBULIN PRODUCTION, METHACRYLATE) BEADS, STATIONARY-PHASE, AFFINITY, LYSOZYME, SEPARATION, PRECONCENTRATION, PURIFICATION, CHROMATOGRAPHY
  • Hacettepe Üniversitesi Adresli: Evet

Özet

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 L-histidine-imprinted poly(hydroxy ethylmethacrylate) based supermacroporous cryogels which can be used for the purification of lysozyme from egg white. A metal chelate monomer [N-methacryloyl-(L)-histidinemethylester (MAH)] forming coordination complex with the template L-histidine in the presence of Cu2+ ions copolymerized with hydroxyethyl methacrylate (HEMA), using N,N'-methylene-bis(acrylamide) (MBAAm) as the cross-linker and ammonium persulfate (APS)/N,N,N',N'-tetramethylene diamine (TEMED) as initiator/activator pair to prepare the MIP cryogel. After that, the template (i.e., L-histidine) was removed using 1 M KSCN solution. The maximum lysozyme adsorption amount was 54.2 mg/g polymer. The relative selectivity coefficients of the MIP cryogel for lysozyme/ribonuclease A and lysozyme/cytochrome c were 4.5 and 2.4 times greater than the non-imprinted poly(HEMA-MAH) (NIP) cryogel, respectively. The resulting MIP cryogels possess excellent long term storage stability and could be used many times without decreasing the adsorption amount significantly. (C) 2011 Elsevier B.V. All rights reserved.