| Summary: | Electrochemical biosensors have potential applications for agriculture, food safety, environmental monitoring, sports medicine, biomedicine, and other fields. One of the primary challenges in this field is the immobilization of biomolecular probes atop a solid substrate material with adequate stability, storage lifetime, and reproducibility. This review summarizes the current state of the art for covalent bonding of biomolecules onto solid substrate materials. Early research focused on the use of Au electrodes, with immobilization of biomolecules through ω-functionalized Au-thiol self-assembled monolayers (SAMs), but stability is usually inadequate due to the weak Au–S bond strength. Other noble substrates such as C, Pt, and Si have also been studied. While their nobility has the advantage of ensuring biocompatibility, it also has the disadvantage of making them relatively unreactive towards covalent bond formation. With the exception of Sn-doped In<sub>2</sub>O<sub>3</sub> (indium tin oxide, ITO), most metal oxides are not electrically conductive enough for use within electrochemical biosensors. Recent research has focused on transition metal dichalcogenides (TMDs) such as MoS<sub>2</sub> and on electrically conductive polymers such as polyaniline, polypyrrole, and polythiophene. In addition, the deposition of functionalized thin films from aryldiazonium cations has attracted significant attention as a substrate-independent method for biofunctionalization.
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