A stable and sensitive non-enzymatic glucose sensor is proposed based on the hybrid thin films of copper on acetanilide/ITO-PET. The electrode was fabricated through a two-step synthesis process including electrochemical deposition of acetanilide on ITO-PET, subsequent by physical vapor deposition of a copper film. The electron microscopy images showed the formation of a porous structure on the ITO-PET sheet. Such modified electrode presents a high sensitivity of 59.95μAmM-1cm-2 with a low limit of detection of 1.501μM. This glucose sensor exhibits a range of linear response between 2 to 12mM. In addition, it includes a variety of advantages as high flexibility, the possibility for scale-up production, low material consumption, low cost, excellent reproducibility, high selectivity, good sensitivity and short response time. Finally, the glucose determination of different human blood serums indicates that the deviation of this sensor from the clinical measurements is less than 5%.