A Fast ImmunosensorBased on Biohybrid Self-AssembledNanostructures for the Detection of KYNA as a Cerebrospinal FluidBiomarker for Alzehimer’s Disease

Although the role of kynurenic acid (KYNA) is not yet fully understood, recent research has implicated this tryptophan (Trp) metabolite as a significant biomarker in neurodegenerative diseases. In this study, we developed an immunosensor platform based on self-assembled polyelectrolyte multilayers (PEMs), employing an enzyme-labeled immunoreagent in a competitive displacement format that requires only a single wash step. This immunosensor enables the detection of KYNA and Trp with detection limits (LOD) of 9 pg/mL and 1.2 ng/mL, respectively. Results validated by traditional ELISA methods indicated elevated levels of KYNA and an increased KYNA/Trp ratio in the cerebrospinal fluid (CSF) of Alzheimer’s patients compared to controls, consistent with previous findings. Additionally, this immunosensor platform can be readily adapted to detect other neuroactive Trp metabolites by substituting specific immunoreagents, supporting a flexible profile-based approach. This platform could serve as a rapid, cost-effective clinical tool for monitoring neurological and psychiatric disorders, potentially advancing therapeutic strategy development.