Herein, a label-free electrochemical aptasensor for alpha-fetoprotein (AFP) analysis was established. The AFP aptamer (AFP-Apt), as the recognition molecule, was immobilized on the surface of a screen-printed carbon electrode, which was modified by gold-platinum metallic nanoparticles and reduced graphene oxide-chitosan-ferrocene nanohybrids (Au-Pt NPs/RGO-CS-Fc), to build the AFP electrochemical aptasensor. The construction process of the aptasensor was characterized by scanning electron microscopy, X-ray photoelectron spectroscopy, Raman spectroscopy, and electrochemical impedance spectroscopy. With the addition of AFP, the formation of the AFP-aptamer conjugation blocked the electron transfer reaction, reducing the differential pulse voltammetric responses of the current of Fc in the RGO-CS-Fc nanohybrids. By optimizing the experimental parameters, AFP could be detected with the dynamic concentration range of 0.001 to 10.0 μg mL-1 and with a detection limit of 0.3013 ng mL-1. In addition, the approach was manifested to have good selectivity, reproducibility, and stability. The fabricated aptasensor had a good recovery rate of 102.36% to 118.09% in real human serum samples. This work demonstrates that the electrochemical aptasensor is a useful tool for analyzing AFP inexpensively, rapidly, and accurately.