Detection of the SARS-CoV-2 nucleoprotein by electrochemical biosensor based on molecularly imprinted polypyrrole formed on self-assembled monolayer.

Herein, we report the development and characterisation of an electrochemical biosensor with a polypyrrole (Ppy)-based molecularly imprinted polymer (MIP) for the serological detection of the recombinant nucleocapsid protein of SARS-CoV-2 (rN). The electrochemical biosensor utilises a Ppy-based MIP formed on a self-assembled monolayer (SAM) at the gold interface to enhance Ppy layer stability on the screen-printed electrode (SPE). Electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV) were employed for the electrochemical characterisation of screen-printed gold electrodes (SPGEs) modified with MIP or non-imprinted polymer (NIP) layers. Removing the rN protein template from the MIP layer increased electron transfer and decreased impedance, indicating the specificity of molecular imprinting. The electrochemical biosensor with a Ppy-based MIP exhibited higher sensitivity than the NIP counterpart, demonstrating its potential for selective rN protein detection. The limit of detection 0.4 nM and 0.2 nM and the limit of quantification 1.3 nM and 0.66 nM values obtained through SWV and EIS, respectively, highlight the biosensor's ability to detect low target protein concentrations. The specificity test confirmed minimal nonspecific binding, reinforcing the reliability of the novel electrochemical sensor with a Ppy-based MIP.