Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

Herein we report the electropolymerization of a scopoletin based molecularly imprinted polymer (MIP) for the detection of lysozyme (Lyz), an enzymatic marker of several diseases in mammalian species. Two different approaches have been used for the imprinting of lysozyme based, respectively, on the use of a monomer-template mixture and on the covalent immobilization of the enzyme prior to polymer synthesis. In the latter case, a multi-step protocol has been exploited with preliminary functionalization of gold electrode with amino groups, via 4-aminothiophenol, followed by reaction with glutaraldehyde, to provide a suitable linker for lysozyme. Each step of surface electrode modification has been followed by cyclic voltammetry and electrochemical impedance spectroscopy, which has been also employed to test the electrochemical responses of the developed MIP. The sensors show good selectivity to Lyz and detect the enzyme at concentrations up to 292 mg/L (20 mu M), but with different performances, depending on the used imprinting approach. An imprinting factor equal to 7.1 and 2.5 and a limit of detection of 0.9 mg/L (62 nM) and 2.1 mg/L (141 nM) have been estimated for MIPs prepared with and without enzyme immobilization, respectively. Competitive rebinding experiment results show that this sensing material is selective for Lyz determination. Tests were performed using synthetic saliva to evaluate the potential application of the sensors in real matrices for clinical purposes.

Automated summary

In this study, electropolymerization of a scopoletin-based molecularly imprinted polymer for lysozyme detection was reported. Two different approaches were used for imprinting lysozyme, and the electrochemical responses of MIPs to Lyz were tested, showing good selectivity and competitive rebinding ability. The MIP prepared with enzyme immobilization exhibited higher imprinting factor and lower detection limit compared to the MIP prepared without enzyme immobilization.