Late state‐of‐the‐art analytical methodologies in chromatography, spectroscopy, and electroanalysis have been developed to meet the challenges of changing environmental and health issues. The modern trends in developing new protocols emphasize economic, portable, nano, or even smaller sample sizes and less time‐consuming processes. This has led to the development of technology‐based biosensors which meet most of the above requirements. The lab‐on‐chip technology exploiting enzyme‐based biosensors has made the analytical processes very efficient, accurate, affordable, and requiring nano‐scale sample sizes. In this review, an attempt is being made to review the literature based on state‐of‐the‐art technology of enzyme‐based biosensors for the detection of biomolecules.