Engineering advanced therapeutic and diagnostic nano‐bio‐platforms (NBPFs) have emerged as rapidly‐developed pathways against a wide range of challenges in antitumor, antipathogen, tissue regeneration, bioimaging, and biosensing applications. Emerged 2D materials have attracted extensive scientific interest as fundamental building blocks or nanostructures among material scientists, chemists, biologists, and doctors due to their advantageous physicochemical and biological properties. This timely review provides a comprehensive summary of creating advanced NBPFs via emerging 2D materials (2D‐NBPFs) with unique insights into the corresponding molecularly restructured microenvironments and biofunctionalities. First, it is focused on an up‐to‐date overview of the synthetic strategies for designing 2D‐NBPFs with a cross‐comparison of their advantages and disadvantages. After that, the recent key achievements are summarized in tuning the biofunctionalities of 2D‐NBPFs via molecularly programmed microenvironments, including physiological stability, biocompatibility, bio‐adhesiveness, specific binding to pathogens, broad‐spectrum pathogen inhibitors, stimuli‐responsive systems, and enzyme‐mimetics. Moreover, the representative therapeutic and diagnostic applications of 2D‐NBPFs are also discussed with detailed disclosure of their critical design principles and parameters. Finally, current challenges and future research directions are also discussed. Overall, this review will provide cutting‐edge and multidisciplinary guidance for accelerating future developments and therapeutic/diagnostic applications of 2D‐NBPFs.