Synthesis of TiO₂ wires was prepared by using two steps method. TiO₂ seed layer was prepared using sol-gel method while TiO₂ wires were grown by using hydrothermal method. The annealing time and temperature during hydrothermal method were varied from 1, 4, and 5 hours and 110 and 120 °C respectively to analyze the effect towards the growth of TiO₂ wires. The morphology of TiO₂ wires formed was characterized using scanning electron microscope (SEM), atomic force microscopy (AFM) and X-ray diffraction (XRD). The electrical characterization of the TiO₂ wire based biosensor was conducted using electrochemical analysis. The cyclic voltammetry response was observed to analyze the performance of the glucose biosensor. The TiO₂ wires have shown a good response towards glucose detection.