There is currently a pressing need to create an electro-analytical approach capable of detecting and monitoring genosensors in a highly sensitive, specific, and selective way. In this work, Functionalized Multiwall Carbon Nanotubes, Graphene, Polypyrrole, and gold nanoparticles nanocomposite (f-MWCNTs-GR-PPy-AuNP) were effectively deposited on the surface of the ITO electrode using a drop-casting process to modify it. The structural, morphological, and optical analysis of the modified ITO electrodes was carried out at room temperature using X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM) images, atomic force microscopy (AFM) and Fourier transform infrared (FTIR) spectra. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to assess the electroanalytical performance of the electrodes after modification. The results showed that using AuNPs and PPy for modification of ITO/f-MWCNTs-GR electrode surfaces is conducive to augmenting the electrochemical performances of the electrodes. ITO/f-MWCNTs-GR showed better results in terms of higher electroactive area formation after modification with PPy and AuNPs. This work aims to figure out how to develop electrochemical biosensors for improved genosensor monitoring.