Advanced nanomaterials for dehydrogenase-based electrochemical sensing: A comprehensive review

Electrochemical biosensors involving dehydrogenase enzymes have attracted significant attention for their capacity to enable sensitive, rapid, and cost-effective detection of therapeutically and environmentally important analytes. This paper provides a comprehensive analysis of the fundamental principles of electrochemical sensing, focusing on dehydrogenase-based systems, including their reaction mechanisms, cofactor dependence, and signal transduction pathways. Conventional detection technologies, such as optical, chromatographic, and mass spectrometry methods, are meticulously evaluated to highlight their limitations in terms of complexity, cost, and applicability to real samples. Recent advances in nanostructured materials, hybrid interfaces, and electrode engineering are systematically evaluated, demonstrating how material selection influences enzyme immobilisation, electron transfer, sensitivity, selectivity, and operational stability. The enzyme–material contact is emphasised as a critical factor influencing sensor functionality. The difficulties of enzyme immobilisation, stability, and interfacial electron transport are meticulously examined to clarify the gap between laboratory performance and real-world applicability. Additionally, issues of material toxicity, environmental impact, and sustainability are taken into account. Ultimately, translational issues such as reproducibility, fouling, scalability, and lack of standardisation are analysed to provide a pragmatic perspective on practical application. This analysis underscores the imperative for systematic interface engineering and standardised evaluation procedures to facilitate the development of reliable dehydrogenase-based electrochemical biosensors for practical applications.