Biosensors are a rapidly expanding field of science owing to their wide variety of applications in healthcare, pharmacology, environmental control, food quality assessment, security and defense, and, most notably, diagnostics. Among various biosensors, electrochemical biosensors are immensely popular due to their high sensitivity, low detection limit, automation competence, low testing cost, and owing to the invention of electrochemical disposable devices capable of dealing with extremely tiny sample quantities. For efficient biosensor development, biomolecule immobilization is a key step which needs the transducer surface to be functionalized. In 2007, polydopamine (PDA) arises as a substrate independent coating material with the capability to functionalize both inorganic and organic surfaces. The chemical structure of PDA enriched with catechol, imine, and amine groups provide an ideal environment for dense immobilization of biomolecules on transducer surface and demonstrates to be simple, accessible, and mild strategy for biosensor design. This review attempts to assemble existing research progressed on PDA-based electrochemical biosensors in terms of enzymatic biosensors (based on H2O2, glucose, alcoholic and laccase), genosensors (DNA sensing), immunosensors and aptasensors. In addition, literature on biosensing of thrombin, tumor indicators, amino acids, and other clinically important analytes has been compiled to offer a thorough review of PDA-based biosensors.