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Abstract: The determination of disease-associated molecules at trace amounts is a key factor for early and efficient diagnosis from human body fluids. Herein, an ultrasensitive electrochemical sensor based on hybrid epitope imprinting and nanomaterial amplification was developed. The hybrid epitope imprinting was achieved by electropolymerization in the presence of two computationally selected and cysteine modified epitopes of neuron specific enolase (NSE), as-synthesized gold nanoparticles (AuNPs), and functional monomer. The AuNPs decorated epitope-mediated hybrid MIPs, as well as the standard hybrid MIPs, were utilized for the preparation of electrochemical sensors to demonstrate the impact of nanomaterial's modification in the polymer network for biomarker sensing. The fabrication process of both sensor types was investigated by employing cyclic voltammetry, square wave voltammetry, atomic force microscopy, and scanning electron microscopy. The biomarker assay using the standard hybrid MIPs resulted in 2.5-fold higher sensitivity compared to single epitope imprints, whereas the AuNP-hybrid MIPs enhanced the sensitivity level to a great extent and allowed the recognition of NSE in human serum in a concentration range of 25-4000 pg/mL. Comparative selectivity studies with non-imprinted polymer resulted in an imprinting factor of 4.2, confirming the high target selectivity of AuNP-MIP cavities. Cross-reaction of the sensor with four reference molecules (dopamine, bovine serum albumin, glucose and elongated peptide) was negligible. As compared to current strategies for epitope imprinting which rely on single epitopes for the formation of molecular cavities, the hybrid epitope-MIPs, particularly with the inclusion of AuNPs have provided more desirable sensing platforms with high sensitivity, affinity and specificity
Template and target information: protein, epitope, neuron specific enolase, NSE
Author keywords: Computationally selected epitopes, dual-template epitope imprinting, AuNP-Decorated imprinted polymer, Electrochemical sensor, Cancer biomarker detection