"Nanobody-based immunodiagnostics: A systematic review of nanobody integration in diagnostics and deep insight into electrochemical immunoassays"

Nanobodies (Nbs) are increasingly recognized as valuable biological molecules with significant potential in the field of diagnostics. Their unique physiochemical characteristics and structural attributes make them ideal candidates for the development of robust diagnostic assays. Nbs have demonstrated efficacy in various diagnostic applications across human, animal, and plant disciplines, meeting the criteria for a reliable assay. This comprehensive review provides an overview of the diverse uses of Nbs in diagnostic assay development and specifically highlights their application in building electrochemical immunosensors. Additionally, it explores various methods employed for the construction of reliable Nb-based biosensors and offers valuable insights and solutions into selecting appropriate platforms through providing practical solutions such as, construct optimization, including framework adjustments, fusion unit introduction, and precise Nb orientation control with a modified cysteine residue. This provides help for researchers in this field to significantly improve performance and reduce the construction time.