Cu2Y2O5 thin films were deposited on non-alkali glass substrates by RF magnetron sputtering. Its crystal structure, microstructure, optical property, mechanical property, and antibacterial activity were investigated by grazing-incidence X-ray diffraction, transmittance spectra, nanoindenter, and antibiotics test, respectively. A single-phase of Cu2Y2O5 was obtained while annealing at 700°C in air and its optical transparency was >80% in the visible region. The hardness and elastic modulus of the film were 6.7GPa and 82GPa, respectively. Antibiotics testing result revealed that Cu2Y2O5 surface had a superior antibacterial performance even at a dark environment. Therefore, Cu2Y2O5 is a promising novel transparent antibacterial hard coating material.