In response to the health risks posed by heavy metal lead, we have developed a self-powered sensor specifically designed for detection of lead ions in this work. This sensor is designed based on an enzyme-free biofuel cell as a self-powered source. The biofuel cell utilizes two-dimensional lamellar Au NPs-Ti3C2Tx heterostructures as the capacitive anode substrate material. The Ti3C2Tx MXene with the excellent charge storage capacity and fast discharge characteristics, provides a large instantaneous current for signal amplification. Furthermore, the DNA walker technique was combined to regulate the loading of ultra-small Au NPs exhibiting glucose oxidase-like properties. Meanwhile, Co, Mn nitrogen-doped carbon-based nanosheets were designed as the cathode with outstanding oxygen reduction reaction catalytic activity. As a result, the constructed enzyme-free biofuel cell has been used to detect Pb2+ with a wide linear range (0.01–7500 nM) and a low detection limit (0.43 pM). It has been successfully applied to the determination of Pb2+ in human plasma, achieving excellent recovery rates (94.0–110.5 %). The developed capacitive anodes not only significantly optimized the performance of the biofuel cell self-powered sensors, but also provided important insights for the future design and construction of biofuel cells.