The development of advanced technologies for the fabrication of functional nanomaterials, nanostructures, and devices has facilitated the development of biosensors for analyses. Two-dimensional (2D) nanomaterials, with unique hierarchical structures, a high surface area, and the ability to be functionalized for target detection at the surface, exhibit high potential for biosensing applications. The electronic properties, mechanical flexibility, and optical, electrochemical, and physical properties of 2D nanomaterials can be easily modulated, enabling the construction of biosensing platforms for the detection of various analytes with targeted recognition, sensitivity, and selectivity. This review provides an overview of the recent advances in 2D nanomaterials and nanostructures used for biosensor and wearable-sensor development for healthcare and healthmonitoring applications. Finally, the advantages of 2D-nanomaterial-based devices and several challenges in their optimal operation have been discussed to facilitate the development of smart highperformance biosensors in the future.