Development of an electrochemical biosensor utilizing a combined aptamer and MIP strategy for the detection of the food allergen lysozyme

This study aims to develop a MIP-Apt-based electrochemical biosensor for the sensitive and selective determination of Lysozyme (Lyz), a food allergen. For the development of the sensor, in the first stage, modifications were made to the screen-printed electrode (SPE) surface with graphene oxide (GO) and gold nanoparticles (AuNPs) to increase conductivity and surface area. The advantages of using aptamer (Apt) and molecularly imprinted polymer (MIP) technology were combined in a single biointerface in the prepared sensing tool. Surface characterization of the biosensor was performed using scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectrometry (XPS), contact angle measurements, cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). A wide linear range from 0.001 to 100 pM was obtained under optimized conditions for the determination of Lyz detection using the proposed MIP-Apt sensing strategy. The limit of detection (LOD) and limit of quantification (LOQ) for Lyz were 3.67 fM and 12 fM, respectively. This biosensor displays high selectivity, repeatability, reproducibility, and long storage stability towards Lyz detection. The results show that a sensitive and selective sensor fabrication is achieved compared with existing methods. Graphical abstract