| Development of MIPs is based on modern affinity technology, which offers the polymer a specific template recognition site in its 3D structure in order to imprint the target molecules. The mechanism of molecularly imprinted polymers (MIPs) and the template is simulated with the relationship of receptor-antibody, enzyme-substrate. MIPs attract biochemists'eyes due to their various advantageous qualities, such as highly specific, chemically stable, high efficient easy preparation, and recyclable. With different purposes, different MIPs have been exploited and applied in many areas, such as biochemical separation, biosensors, mimic enzyme catalysts, and clinical analysis.Acrylamide(AM) and methacrylic acid(MAA) were used as functional monomer, and ethylene glycol dimethacrylate, azodiisobutyronitrile, dimethylsulfoxide were used respectively as cross-linker, initiator and porogen mateial to synthesize Molecularly Imprinted Polymers (MIPs) by bulk polymerization, for adsorption of glutathione (GSH), and the polymers were named as GSH-MIPAMs and GSH-MIPMAAs. The research of physical and chemical properties of polymers revealed GSH-MIPs with specific three-dimensional network structure and the corresponding functional groups.Evaluation of various polymers by static experiment indicated the optimum molar ratio of template GSH to functional monomer AM and MAA was 1:5 and 1:4 respectively, and the polymers prepared with the optimal molar ratio were the subjects of the follow-up tests. The influences of the pH of loading buffer, adsorption equilibrium time, the concentration of loading buffer on the adsorption of GSH-MIPs were investigated. The results of showed that the optimum pH of loading buffer were 3.00 and 5.00 to GSH-MIPAMs and GSH-MIPMAAs respectively, the optimal concentration of loading buffer were 1.50-2.00g/L to GSH-MIPAMs, and 1.00-1.50g/L to GSH-MIPMAAs, the adsorption equilibruium time of GSH-MIPAMs and GSH-MIPMAAs were 20h and 18h, respectively. By Scatchard analysis, the GSH binding site was found in GSH-MIPs. And Kd of GSH-MIPAMs and GSH-MIPMAAs were 2.11×10-3μmol/L and 5.13×10-4μmol/L, Qmax of GSH-MIPAMs and GSH-MIPMAAs were 39.24μmol/g and 102.16μmol/g. In addition, to gain better results on the selectivity of GSH-MIPs, besides GSH, the analogues GSSG, Gly, Glu and Cys were also tested, and the results showed that GSH-MIPs had low adsorption for the analogues.The method for extraction of GSH from yeast cells samples was validated based on the above experiments. At the optimal condition, the maximum extraction capacity of GSH-MIPAMs and GSH-MIPMAAs were found to be 24.76μmol/g and 46.91μmol/g, and the extraction efficiency were 41% and 77%, respectively, proved to be another option for purification of GSH.
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