A novel electrochemical sensor was fabricated for the investigation of the effect of electron transfer between glutathione (GS) protected gold nanoparticles (AuNPs) and a new C 4 -thiol functionalized viologen axle molecule (V 2+ -SH) on pseudorotaxane formation. The sensor was fully screen printed with silver (Ag) ink on a flexible polyethylene terephthalate (PET) substrate. Square wave voltammetry (SQWV) based response of the sensor toward V 2+ -SH in the presence of Au 25 or 4nm-Au demonstrated one-electron transfer processes, based on 59mV changes in peak potentials to more negative values. The response demonstrated the capability of the novel electrochemical sensor to be used for studying complex systems in volumes as small as 5–10μL, with less sample preparation time.