Detection and quantification of diverse analytes such as molecules, cells receptor and even particles and nanoparticles, play an important role in biomedical research, particularly in electrochemical sensing platform technologies. In this study, gold nanoparticles (AuNPs) prepared by green synthesis from Sargassum sp. were characterized using ultraviolet-visible (UV-Vis) and Fourier transform-infrared (FT-IR) spectroscopies, X-ray diffraction (XRD), scanning electron microscopy (SEM), dynamic light scattering (DLS) and zeta potential (ζ) obtaining organic capped face-centered cubic 80–100 nm AuNPs with an excellent stability in a wide range of pH. The AuNPs were used to modify a carbon nanotubes-screen printed electrode (CNT-SPE), through the drop-casting method, to assemble a novel portable electrochemical sensing platform for glucose, using a novel combination of components, which together have not been employed. The ability to sense and measure glucose was demonstrated, and its electrochemical fundamentals was studied using cyclic voltammetry (CV). The limits of detection (LOD) and quantification (LOQ) to glucose were 50 μM and 98 μM, respectively, and these were compared to those of other sensing platforms.