MXene emerged as decent 2D material and has been exploited for numerous applications in the last decade. The remunerations of the ideal metallic conductivity, optical absorbance, mechanical stability, higher heterogeneous electron transfer rate, and good redox capability have made MXene a potential candidate for biosensing applications. The hydrophilic nature, biocompatibility, antifouling, and anti‐toxicity properties have opened avenues for MXene to perform in vitro and in vivo analysis. In this review, the concept, operating principle, detailed mechanism, and characteristic properties are comprehensively assessed and compiled along with breakthroughs in MXene fabrication and conjugation strategies for the development of unique electrochemical and optical biosensors. Further, the current challenges are summarized and suggested future aspects. This review article is believed to shed some light on the development of MXene for biosensing and will open new opportunities for the future advanced translational application of MXene bioassays.