One of the most important factors for the proper functioning of enzymatic electrochemical biosensors is the enzyme immobilization strategy. In this work, glucose oxidase was covalently immobilized using pentafluorophenyl methacrylate (PFM) by applying two different surface modification techniques (plasma polymerization and plasma-grafting). The grafted surface was specifically designed to covalently anchor enzyme molecules. It was observed using QCM-D measurements the PFM plasma-grafted surfaces were able to retain a higher number of active enzyme molecules than the PFM polymerized surfaces. An amperometric glucose biosensor using titanium dioxide nanotubes array (TiONTAs) modified by PFM plasma-grafted surface was prepared. The resulting biosensor exhibited a fast response and short analysis time (approximately eight minutes per sample). Moreover, this biosensor achieved high sensitivity (9.76 μA mM) with a linear range from 0.25 to 1.49 mM and a limit of detection (LOD) equal to 0.10 mM of glucose. In addition, the glucose content of 16 different food samples was successfully measured using the developed biosensor. The obtained results were compared with the respective HPLC value and a deviation smaller than 10% was obtained in all the cases. Therefore, the biosensor was able to overcome all possible interferences in the selected samples/matrices.