Nanotechnology-enabled biosensors are reviewed in this article, focusing on their fundamentals, materials, applications, challenges, and future scope. Traditional sensors used to measure chemical, physical, or biological factors were large, inaccurate, and required manual interpretation. Nanomaterials such as nanoparticles, nanowires, carbon nanotubes (CNTs), nanorods, and quantum dots (QDs) are commonly used in nanobiosensors. These materials offer high sensitivity, large surface area, high stability, and high thermal and electrical conductivity. Nanobiosensors can detect a wide range of clinically important molecules, including nucleic acids, viruses, bacteria, cancer antigens, pharmaceuticals, narcotics, toxins, contaminants, and even whole cells in various sensing media. Recent developments in nanomaterials have significant potential for point-of-care diagnostics and early disease screening. This review discusses the principles of nanotechnology-enabled biosensors and their architectural design, including the classification of nanobiosensors, various attributes, and fabrication methods. The review also broadens perspectives on the materials used in biosensors and the potential applications of nanotechnology-enabled biosensors. Biosensors are defined as devices that detect and quantify analytes using specific biochemical reactions. They are classified based on bioreceptors, transducers, technology, and detection systems. Different classifications of biosensors are illustrated in the article. Biosensors can be classified into four categories based on bioreceptors: aptamer or nucleic acid-based biosensors, enzymatic biosensors, immunosensors, and microbial or whole-cell biosensors. Based on transducers, biosensors are classified into electrochemical, electronic, thermal, optical, and mass-based or gravimetric. Challenges in biosensor commercialization include biomaterial presence, sensor device development, and system integration. High cost is a significant obstacle to mass fabrication. Table 1 illustrates some advantages and disadvantages of biosensors. Nanomaterials are incorporated into biosensors to enhance sensitivity, catalysis, low-potential reactivity, and electron transfer from the active reaction center to the electrode surface.Nanotechnology-enabled biosensors are reviewed in this article, focusing on their fundamentals, materials, applications, challenges, and future scope. Traditional sensors used to measure chemical, physical, or biological factors were large, inaccurate, and required manual interpretation. Nanomaterials such as nanoparticles, nanowires, carbon nanotubes (CNTs), nanorods, and quantum dots (QDs) are commonly used in nanobiosensors. These materials offer high sensitivity, large surface area, high stability, and high thermal and electrical conductivity. Nanobiosensors can detect a wide range of clinically important molecules, including nucleic acids, viruses, bacteria, cancer antigens, pharmaceuticals, narcotics, toxins, contaminants, and even whole cells in various sensing media. Recent developments in nanomaterials have significant potential for point-of-care diagnostics and early disease screening. This review discusses the principles of nanotechnology-enabled biosensors and their architectural design, including the classification of nanobiosensors, various attributes, and fabrication methods. The review also broadens perspectives on the materials used in biosensors and the potential applications of nanotechnology-enabled biosensors. Biosensors are defined as devices that detect and quantify analytes using specific biochemical reactions. They are classified based on bioreceptors, transducers, technology, and detection systems. Different classifications of biosensors are illustrated in the article. Biosensors can be classified into four categories based on bioreceptors: aptamer or nucleic acid-based biosensors, enzymatic biosensors, immunosensors, and microbial or whole-cell biosensors. Based on transducers, biosensors are classified into electrochemical, electronic, thermal, optical, and mass-based or gravimetric. Challenges in biosensor commercialization include biomaterial presence, sensor device development, and system integration. High cost is a significant obstacle to mass fabrication. Table 1 illustrates some advantages and disadvantages of biosensors. Nanomaterials are incorporated into biosensors to enhance sensitivity, catalysis, low-potential reactivity, and electron transfer from the active reaction center to the electrode surface.