A nanoporous gold nanosphere (pAu NS) was synthesized to load high-content glucose oxidase (GOx) and ferrocene (Fc) for the successful preparation of a new gold nanoprobe. After the specific recognition of the tumor biomarker of carcinoembryonic antigen (CEA) at a gold electrode based aptasensor, this GOx and Fc dually functionalized pAu NS nanoprobe was further used for sandwich immunoreaction and signal tracing. Based on the Fc-mediated GOx-catalytic reaction, the gold nanoprobes quantitatively captured onto the electrode surface produced a sensitive electrochemical signal corresponding to the protein recognition events, which led to the development of a new biosensing method for CEA measurement. Both the high loading of GOx and Fc on the pAu NS nanocarrier and the enzymatically catalytic reaction of the nanoprobe greatly amplify the electrochemical signal; meanwhile, the immobilization of the Fc mediator on this enzyme nanoprobe and the highly specific aptamer recognition drastically decrease the background current, resulting in the achievement of ultrahigh sensitivity of the method. Under optimum conditions, this method shows an excellent analytical performance including a wide linear relationship of five-order of magnitude and a low detection limit down to 0.45 pg mL(-1). Thus this pAu NS based gold nanoprobe and the proposed immunoassay method provide great potential for practical applications.