The release of heavy metalHeavy metals ionsEnvironmental analysis, chemicalEnvironmental toxins, and pesticide residues to the environmentEnvironment is a matter of global concern. Conventional techniques such as chromatography, spectrophotometry, and immunoassays, often involve complex protocols, costly instrumentation and skilled personnel. These limitations cause hindrance towards quick response in analysis and monitoringMonitoring of dynamic environmentalEnvironmental conditions. The rapid advancements in nanotechnologyNanotechnology have led to the emergence of new class of sensors called nanobiosensorsNanobiosensor, which integrate the specificitySpecificity of biological recognition elements with the exceptional properties of nanomaterials. These sensors are revolutionizing the landscape of environmental analysisEnvironmental analysis and monitoringMonitoring by providing tools capable of detecting environmentalEnvironmental pollutantsPollutants, screening groundwater, pathogens, and to monitor other toxic substances at trace levels. This chapter delves into the principles of nanobiosensorsNanobiosensor, their design and working, highlighting the broad spectrum of their applications in environmental analysisEnvironmental analysis and monitoringMonitoring.