In this study, we demonstrated a one-pot synthesis of Cu nanoplates (Cu NPs) with high purity and ‘‘clean surface”. Different from surfactant-assisted method, we tailored the architecture by controlling the ripening time of Cu seeds. Citrate is introduced as a structure-directing agent. Since the ‘‘clean surface” provide more available active sites, the as-prepared Cu NPs show a superior performance when utilized in the non-enzymatic biosensors. In the detection of nitrite, the sensor constructed by Cu NPs exhibited a superior sensitivity (303.1 ± 10.18 μA·mM-1·cm-2), and the detection range is 5.0 mM. On the contrary, sensitivity for the Cu NPs by surfactant-assisted method is negligible. As applied in the glucose sensors, detection range by Cu NPs could reach 36.0 mM. And the sensitivity under ambient condition (298 K) is 561.93 ± 3.72 μA·mM-1·cm-2, which is ca. 2.11-folds higher than that for Cu NPs by surfactantassisted method. In the detection of H2O2, detection range is 10.0 mM, and the enhancement factor for detection range is up to 2.0-folds. The superior performance can also be extended to the catalysis for the reduction of 4-nitro-phenol. Since no additional noble metals was applied, this strategy would open up a new avenue for utilizing non-noble metals.