Innovative Fatty Acid-Guided Biosensor Design for Neutrophil Gelatinase, a Prognostic and Diagnostic Biomarker for Chronic Kidney Disease

Chronic kidney disease (CKD) afflicts 850 million people worldwide, with an estimate that it is the 5th highest cause of years of life lost (YLLs). Standard confirmatory procedures for disease are blood and urine analysis with ultrasound for confirmation. Neutrophil gelatinase-associated lipocalin (NGAL) has been established as a prognostic biomarker, especially for the pre-clinical stages of CKD, thus presenting itself as a dependable predictor of the progression. With the aim of designing diagnostics, fatty acids were explored as potential biorecognition elements for the selective capture of NGAL. Three fatty acids-linoleic acid, arachidonic acid, and retinoic acid-were shortlisted as plausible candidates based on their known affinity toward lipocalin family proteins. Docking followed by molecular dynamics simulations were employed to evaluate the binding affinity and stability of each complex. Among them, linoleic acid exhibited the most favorable interaction, as evidenced by the lowest binding free energy. Subsequently, fluorescence and electrochemical techniques-square-wave voltammetry, differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectroscopy (EIS)-were systematically compared for qualitative and quantitative checking of the accuracy of NGAL detection. Amongst the electrochemical techniques, differential pulse voltammetry DPV demonstrated superior analytical performance with an LOD of 0.05 ng/mL with a sensitivity of 23.2 µA/cm²/pg. To the best of our knowledge, this is the first report of a fatty acid-based biosensor platform for NGAL detection, presenting a novel approach for CKD diagnostics. The sensitivity obtained is comparable with available NGAL detection methods yet cost-effective and robust.