Creating a macromolecular receptor by affinity imprinting

A molecularly imprinted polymeric receptor for trypsin was synthesized by employing a novel technique that is a combination of affinity separation and molecular imprinting. An enzyme–inhibitor complex of trypsin and N‐acryloyl para‐aminobenzamidine was polymerized with acrylamide and N,N'‐methylene bis‐acrylamide. Template trypsin was extracted out to obtain an affinity‐imprinted polymer. Control experiments were performed to demonstrate the synergistic affinity‐imprinting effect. The percentage of crosslinker used was the crucial factor in determining the imprinting efficacy of the polymers. Imprinted polymer containing 50% crosslinker exhibited a linear Scatchard plot. Unlike non‐imprinted gel, the receptor exhibited almost exclusive recognition of trypsin in an individual batch experiment as well as from a mixture of trypsin and chymotrypsin. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 1075–1083, 2001