The application of biofuel cells (BFCs) for the construction of self‐powered electrochemical biosensors has recently received enormous attention with exciting advancements. The principle of BFCs for electrochemical biosensing is that the BFCs’ power output which is directly extracted by the biological reactions with the help of biocatalysts is closely related to the analyte's concentration. Such unique feature makes BFCs based electrochemical biosensors with two‐electrode system to be operated without the need of any external power source, which provides a more effective way for the miniaturization and convenient construction of electrochemical biosensors than the traditional electrochemical biosensors with three‐electrode system. Following our previous reviews (Electroanalysis 2012, 24, 197–209; Electroanalysis 2015, 27, 1786–1801), the current review is focused on the recent development and advances in the construction of BFCs based self‐powered electrochemical biosensors which are reported in the years between 2015 and 2018, with a particular emphasis on their possible practical applications in the fields of medical diagnosis, environmental monitoring, food analysis and wearable devices. For these reported BFCs‐based self‐powered electrochemical biosensors, the design approaches are still within the scope of effects (i.e., substrate, inhibition, blocking, gene regulation, enzyme, co‐stabilization, competition, and hybrid effects) summarized by our previous reviews. The outlook, challenges and developing tendency of the future research for the construction of BFCs based self‐powered electrochemical biosensor are discussed in the end. We expect that the advanced electrochemical biosensors based on BFCs with the unique self‐powered capability will continue attracting increasing research interest and lead to new opportunities in various attractive applications related to analytical chemistry.