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Abstract: A facile, general, and highly efficient approach to prepare uniform core-shell molecularly imprinted polymer (MIP) particles with enzyme inhibition potency is described for the first time, which involves the combined use of molecular imprinting and controlled/"living" radical polymerization (CRP) techniques as well as surface-anchoring strategy. The thickness of the enzyme-imprinted surface layers of the core-shell MIP microspheres had a significant influence on their binding properties, and only those with their thickness comparable with the diameters of the targeted enzymes could afford enzyme-MIPs with optimal specific bindings. The as-prepared enzyme-MIPs were found to have homogeneous binding sites and high template binding capacities, affinity, and selectivity, and they proved to show much higher enzyme inhibition potency than the small inhibitor by 3 orders of magnitude (i.e., the enzyme inhibition constant of every binding site of the MIP microspheres was about one-thousandth of that of the small inhibitor), mainly due to the formation of strong long-range secondary interactions between enzymes and imprinted pockets. In addition, the general applicability of our strategy was confirmed
Template and target information: trypsin