Electrochemical evaluation of a protein-imprinted polymer receptor

The selective detection and quantification of macromolecular targets is a fundamental biological mechanism in nature. Molecularly imprinted polymers (MIPs) have been identified as one of the most promising synthetic alternatives to bioreceptors. However, expanding this methodology towards selective recognition of bulky templates such as proteins appears to be extremely challenging due to problems associated with removal of the template from the polymeric network. In this study, polymer imprinted with troponin T (TnT) was assessed using electrochemical methods and the influence of various extraction methods, including conventional immersion extraction, thermal annealing and ultrasonic-assisted extraction, on the binding characteristics of the troponin-to-imprinted polymer receptor was elucidated. Cyclic voltammetric deposition of o-phenylenediamine (o-PD) film in the presence of TnT as a template was performed in acetate buffer (0.5 M, pH 5.2) on a gold substrate. Solvent extraction of the target molecule was optimised and followed by subsequent washing with water. The electrochemistry of a ferro/ferricyanide probe was used to characterise the TnT MIP receptor film. The incubation of the TnT MIP receptor-modified electrode with respect to TnT concentration resulted in a suppression of the ferro/ferricyanide redox current. The dissociation constant (K D) was calculated using a two-site model of template affinity for the TnT MIP receptor. The synthetic TnT MIP receptor had high affinity for TnT with a K D , of 2.3 × 10-13 M.