PANI/CMWCNTs immunosensor with KNN-Ridge for high-throughput COX-2 screening

Chronic inflammation is tightly linked to atherosclerosis and malignancies. Cyclooxygenase-2 (COX-2) is a pivotal target for anti-inflammatory drug discovery. However, conventional screening approaches for natural products are limited by low throughput, insufficient precision, and prolonged assay durations. Herein, a high-performance COX-2-targeted electrochemical immunosensor was constructed with polyaniline/carboxylated multiwalled carbon nanotube (PANI/CMWCNTs) nanocomposite as conductive sensing interface. Benefiting from the synergistic electron-transport network of dual nanomaterials, this sensor delivers distinct quantitative advantages over traditional single-carbon-material and bare glassy carbon electrode sensors, including greatly boosted detection sensitivity and superior signal stability. Fabrication parameters were systematically optimized using the Box-Behnken response surface methodology coupled with cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under optimized conditions, the developed immunosensor exhibited a low limit of detection (LOD) of 76 pg/mL. By By combining electrochemical response signals with K-nearest neighbor-ridge regression (KNN-Ridge) machine learning, an intelligent screening platform was further built. The integrated system achieves 60% classification accuracy for discriminating herbal active ingredients, with a Pearson correlation coefficient of 0.71 for concentration trends). This approach captures the rough variation trend of analyte concentration, significantly shortening screening duration and meeting the demand for qualitative preliminary screening that traditional assays cannot satisfy. This work establishes an integrated "sensing-intelligent trend analysis" strategy for rapid, preliminary screening of COX-2-targeted anti-inflammatory components from natural products, facilitating intelligent auxiliary analysis and traceability assessment of bioactive constituents in traditional herbal medicines.