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.
Bereli
Nilay
N
Department of Chemistry, Biochemistry Division, Hacettepe University, Ankara, Turkey.
Andaç
Müge
M
Baydemir
Gözde
G
Say
Ridvan
R
Galaev
Igor Yu
IY
Denizli
Adil
A
D000327:Adsorption; D002021:Buffers; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels; D006863:Hydrogen-Ion Concentration; D008855:Microscopy, Electron, Scanning; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity
D002021:Buffers; D005782:Gels; D011506:Proteins