In medicine and biotechnology, traditional in vitro diagnostics require trained personnel in centralized laboratories to perform time‐consuming experiments with costly, large, and bulky devices. Therefore, the development of highly sensitive biosensor devices is essential for successful bioanalytical applications. Biosensors are based on the coupling of a biorecognition element that is responsible for the specific recognition of the analytes of interest and a physicochemical transducer that converts the chemical signal into an electrical output signal. This electrical output signal is then processed and transferred to a display by the electronic system. In addition, biosensors have become highly versatile platforms for a broad range of applications in different research areas because of their ease of use and capability to operate in complex media. The advancements made in micro‐ and nanoscale fabrication have enabled the integration of biological and/or chemical species with microelectronics to result in the mass production of biochips in a cost‐effective manner. In the medical diagnostic field, biosensors and biochips continue to play a critical role that leads to effective clinical outcomes and promotes general public health by enabling rapid diagnosis of diseases in the early stages. In recent years, significant research has been conducted on the design and fabrication of biosensors for the detection of various biomarkers of diseases by taking advantage of the various biosensor features, including sensitivity, selectivity, low cost, and rapid response time. In this article, focus is placed on the principles of operation, transduction, and immobilization mechanisms of biosensors, and the techniques and materials used for the fabrication of biosensors with emphasis placed on the most commonly reported electrochemical biosensors. The nanoscale electrode structures that have gained great interest for enzyme immobilization are introduced, and an overview into the development of novel, sophisticated, and miniaturized self‐powered biosensor system is discussed.