▎ 摘　　要

We described a sensitive, label-free electrochemical immunosensor for the detection of carcinoembryonic antigen. It is based on the use of a glassy carbon electrode (GCE) modified with a multi-layer films made from Prussian Blue (PB), graphene and carbon nanotubes by electrodeposition and assembling techniques. Gold nanoparticles were electrostatically absorbed on the surface of the film and used for the immobilization of antibody, while PB acts as signaling molecule. The stepwise assembly process was investigated by differential pulse voltammetry and scanning electron microscopy. It is found that the formation of antibody-antigen complexes partially inhibits the electron transfer of PB and decreased its peak current. Under the optimal conditions, the decrease of intensity of the peak current of PB is linearly related to the concentration of carcinoembryonic antigen in two ranges (0.2-1.0, and 1.0-40.0 ng center dot mL(-1)), with a detection limit of 60 pg center dot mL(-1) (S/N = 3). The immunosensor was applied to analyze five clinical samples, and the results obtained were in agreement with clinical data. In addition, the immunosensor exhibited good precision, acceptable stability and reproducibility.