Reagentless and sensitive detection of tumor biomarkers using label-free electrochemical immunosensors is highly desirable for early and effective cancer diagnosis. Herein, we present a label-free electrochemical immunoassay platform based on surface-confined Prussian blue (PB) redox probes for sensitive and reagentless determination of carcinoembryonic antigen (CEA). To facilitate the electron transfer of probes and improve sensitivity, Au nanoparticles and PB (Au-PB) are electrochemically co-deposited on a carbon nanotube (CNT) modified glassy carbon electrode (GCE). A polydopamine (pDA) layer is coated on the Au-PB nanocomposite layer in situ as a bifunctional linker. In addition to improving the stability of PB, pDA also provides reducibility for the preparation of gold nanoparticles, which offers an interface for anti-CEA antibody immobilization. The fabricated immunosensor has good stability and is able to reagentlessly detect CEA over a wide range (0.005-50 ng mL-1) with high reproducibility. Furthermore, the immunosensor was used for determination of CEA in human serum samples.This journal is © The Royal Society of Chemistry.