Background: The electrochemical biosensor is one of the typical sensing devices based on transducing the biochemical events to electrical signals. In this type of sensor, an electrode is a key component that is employed as a solid support for immobilization of biomolecules and electron movement. Thanks to numerous nanomaterials that possess the large surface area, synergic effects are enabled by improving loading capacity and the mass transport of reactants for achieving high performance in terms of analytical sensitivity. Main body: We categorized the current electrochemical biosensors into two groups, carbon-based (carbon nanotubes and graphene) and non-carbon-based nanomaterials (metallic and silica nanoparticles, nanowire, and indium tin oxide, organic materials). The carbon allotropes can be employed as an electrode and supporting scaffolds due to their large active surface area as well as an effective electron transfer rate. We also discussed the non-carbon nanomaterials that are used as alternative supporting components of the electrode for improving the electrochemical properties of biosensors. Conclusion: Although several functional nanomaterials have provided the innovative solid substrate for high performances, developing on-site version of biosensor that meets enough sensitivity along with high reproducibility still remains a challenge. In particular, the matrix interference from real samples which seriously affects the biomolecular interaction still remains the most critical issues that need to be solved for practical aspect in the electrochemical biosensor. Recently, various electrochemistry-driven biosensing methods have been introduced for simple and miniaturized analytical devices for on-site analysis. This trend can be applied to replace the commercial lab instruments manufactured by the renowned in vitro diagnosis