Preparation of surface molecularly imprinted polymeric microspheres and their recognition property for basic protein lysozyme.
The surface imprinting of basic protein lysozyme (Lys) was carried out by designing a new route. The copolymerization of N-vinylpyrrolidone (NVP) and 2-hydroxyethyl methacrylate (HEMA) was first conducted in an inverse suspension polymerization system, and the crosslinked copolymeric microspheres HEMA/NVP were prepared. Subsequently, the esterification reaction of methacryloyl (MAO) chloride with the hydroxyl groups on the surfaces of HEMA/NVP microspheres was performed, and the modified microspheres MAO-HEMA/NVP, on which a mass of polymerisable double bonds were introduced, were obtained. In the presence of lysozyme, by initiating of K(2)S(2)O(8)-NaHSO(3), the monomer methacrylic acid (MAA) in the solution was crosslink-polymerized on the surfaces of MAO-HEMA/NVP microspheres, resulting in the surface imprinting of lysozyme. After removing the template molecules, the lysozyme molecule-surface-imprinted material MIP-HEMA/NVP was obtained. Because there were strong interactions between lysozyme and monomer MAA, electrostatic interaction and hydrogen bonding, the lysozyme molecule-surface imprinting was successfully realized. The MIP-HEMA/NVP microspheres have very high binding affinity for lysozyme, and the binding capacity gets up to 216 mg/g. It is more important that MIP-HEMA/NVP microspheres have specific recognition selectivity for lysozyme, and the selectivity coefficient for lysozyme with respect to bovine hemoglobin (BHb), which was used as a contrast protein in the experiments, actually reaches 31.07. In the respect of protein imprinting, the imprinting material with such high performance is unwonted.
Gao
Baojiao
B
Department of Chemical Engineering, North University of China, Taiyuan 030051, People' s Republic of China. gaobaojiao@126.com
Fu
Hongyan
H
Li
Yanbin
Y
Du
Ruikui
R
D000327:Adsorption; D000818:Animals; D002417:Cattle; D006454:Hemoglobins; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008689:Methacrylates / Q000737:chemistry*; D008863:Microspheres; D054802:Molecular Imprinting / Q000379:methods*; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D011760:Pyrrolidinones / Q000737:chemistry*; D013055:Spectrophotometry, Infrared; D013696:Temperature
D006454:Hemoglobins; D008689:Methacrylates; D011760:Pyrrolidinones; C005044:hydroxyethyl methacrylate; C042670:N-vinyl-2-pyrrolidinone; D009113:Muramidase