Hereby, one type of facile-green copper nanoclusters (CuNCs) derived from ovalbumin (CuNCs@OVA) has been innovatively synthesized with a quantum yield of 5.8%. Meaningfully, this nanoclusters exhibited the multiple advantages including satisfactory photostability, low-toxicity and satisfactory stability. By the characterizations in detail, there existed –COOH, –OH, –NH2 and C˭O groups on their surfaces of the prepared CuNCs@OVA, providing their possible applications for sensing. Significantly, the fluorescence of the as-prepared CuNCs@OVA could be quenched by introducing Vitamin B1, while they exhibited the enhanced fluorescence by adding into doxycycline. On the basis of both the fluorescence quenching and enhancement, CuNCs were further employed as the fluorescent probe for assaying both Vitamin B1 and doxycycline. Consequently, it displayed a linear range of 1–1000nM with the detection limit of 380pM for assaying VB1, as well as a linear range from 1 to 1000μM with the detection limit of 270nM for detecting doxycycline, which indicates the potential applications of CuNCs@OVA for the actual-sample detections.