In this paper, we first synthesis three-dimensional jasmine-like Cu@L-aspartic acid(L-ASP) inorganic-organic hybrid nanoflowers to load palladium-platinum nanoparticles (Pd-Pt NPs) as the signal enhancer in order to quantify intracellular speckle-type POZ domain protein. Scanning electron microscope, fourier transform infrared, energy dispersive spectrometer, X-ray photoelectron spectroscopy analysis was used to characterize the newly synthesized materials. The newly formed Cu@L-Asp/Pd-PtNPs can catalyze the decomposition of hydrogen peroxide and exhibit excellent catalytic performance. When different concentration of speckle-type POZ domain protein is captured by speckle-type POZ domain protein antibody linked to the surface of Cu@L-Asp/Pd-Pt NPs, the current signal decreases with the increase concentration of speckle-type POZ domain protein. After optimization, the speckle-type POZ domain protein immunosensor exhibited a good linear response over a concentration range from 0.1-1 ng mL-1 with a low detection limit of 19 fg mL-1. The proposed sensor demonstrates good stability within 28 days, acceptable reproducibility (RSD = 0.52%) and selectivity to the speckle-type POZ domain protein in the presence of possible interfering substances and has potential application for detecting other intracellular macromolecular substances.