Development of a novel high-affinity anti-NGAL monoclonal antibody pair for sensitive detection of acute kidney injury.

Neutrophil Gelatinase-Associated Lipocalin (NGAL) is an early marker for Acute Kidney Injury (AKI) recommended by the International Society of Nephrology (ISN). This study obtained recombinant NGAL protein via a prokaryotic expression system. After immunizing BALB/c mice, four NGAL-specific monoclonal antibodies were screened using hybridoma technology. Through paired screening, mAb-14/mAb-21 was identified as the optimal antibody pair. Based on this, two immunological detection methods were preliminarily established: magnetic particle chemiluminescence assay (MPs-CLIA) and colloidal gold immunochromatography assay (GICA). According to the performance evaluation, MPs-CLIA exhibited a detection limit of 0.53 ng/mL across a range of 15.625-500 ng/mL, with a superb fit (R2 = 0.9996). It demonstrated a significant association with the commercial kit (R2 = 0.9489, rs = 0.9499, P < 0.0001). The recovery rate for spiked samples ranged from 92.08% to 92.33% (CV < 10%). In clinical serum samples, GICA obtained 97.5% accuracy and detected analytes down to 12.5 ng/mL. The established detection system addresses diverse application needs, ranging from highly sensitive quantification to rapid screening, and provides a reliable technical solution for the early diagnosis of AKI.