Surface-grafted molecularly imprinted polymers for protein recognition

A technique for coating microplate wells with molecularly imprinted polymer s (MIP's) specific for proteins is presented. 3-Aminophenylboronic acid was polymerized in the presence of the following templates: microperoxidase, h orseradish peroxidase, lactoperoxidase, and hemoglobin, via oxidation of th e monomer by ammonium persulfate. This process resulted in the grafting of a thin polymer layer to the polystyrene surface of the microplates. Imprint ing resulted in an increased affinity of the polymer toward the correspondi ng templates. The influence of the washing procedure, template concentratio n, and buffer pH on the polymer affinity was analyzed. It was shown that th e stabilizing function of the support and spatial orientation of the polyme r chains and template functional groups are the major factors affecting the imprint formation and template recognition. Easy preparation of the MIPs, their high stability, and their ability to recognize small and large protei ns, as well as to discriminate molecules with small variations in charge, m ake this approach attractive and broadly applicable in biotechnology, assay s and sensors.