Graphdiyne (GDY), a new kind of two-dimensional carbon nanomaterial, has abundant chemical bonds, a highly πconjugated structure, uniformly distributed pores, a large surface area, and good biocompatibility and has been applied in drug delivery, biosensing, antibacterial processes, and cancer therapy. Herein, this review is intended to provide an introduction of current advances in bioapplications of GDY, which covers the synthetic methods, structure and properties of GDY, and its application in antimicrobial processes, cancer therapy, biosensing, and bioimaging. Finally, we provide a perspective on future development in research relevant to GDY. We believe that this review can provide meaningful assistance for breakthroughs and innovations in the field of GDY.