In this work, a new network nanocomposite composed of polypyrrole hydrogel (PPy hydrogel) loaded gold nanoparticles (AuNPs) was prepared. The PPy hydrogel was directly synthesized by mixing the pyrrole monomer and phytic acid, and the mixed solution can be gelated to form hydrogel at once. The three-dimensional network nanostructured PPy hydrogel not only provided a greater effective surface area for increasing the quantity of immobilized biomolecules and facilitated the transport of electrons and ions, but also exhibited an improved conductivity. Meanwhile, the electrodeposited AuNPs on the PPy hydrogel can further increase the specific surface area to capture a large amount of antibodies as well as improve the capability of electron transfer. The network PPy hydrogel/Au nanocomposites were successfully employed for the fabrication of a sensitive label-free amperometric immunosensor. Carcinoembryonic antigen (CEA) was used as a model protein. The proposed immunosensor exhibited a wide linear detection range from 1 fg mL(-1) to 200 ng mL(-1), and an ultralow limit of detection of 0.16 g mL(-1) (S/N = 3), and it also possessed good selectivity. Moreover, the detection of CEA in ten human serums showed satisfactory accuracy compared with the data determined by ELISA, indicating that the immunosensor provided potential application for clinical diagnosis.