Abstract
Paper-based lateral flow assays (LFAs) are among the most widely used biosensing platforms for point-of-care testing (POCT). However, the conventional colloidal gold label of LFAs show low sensitivity and limited quantitative capacity. Alternatively, the use of enzyme/chemical reaction-based signal amplification with structural modifications has enhanced analytical capacity but requires multiple user interventions as a trade-off, increasing complexity, test imprecision, and time. These platforms are also difficult to manufacture, limiting their practical applications. In this study, within the current LFA production framework, we developed a highly sensitive, automated, universal, and manufacturable LFA biosensing platform by (i) incorporating gold nanoparticles into a polymer-networked peroxidase with an antibody as a new scheme for enhanced enzyme conjugation and (ii) integrating a mass-producible and time-programmable amplification part based on a water-swellable polymer for automating the sequential reactions in the immunoassay and signal amplification, without compromising performance, simplicity, and production feasibility. We applied this platform to evaluate cardiac troponin I (cTnI), a gold-standard biomarker for myocardial infarction diagnosis. Quantitative analysis of cTnI in clinical setting remains limited to the laboratory-based high-end and costly standard equipment. Coupled with an enzyme-catalyzed chemiluminescence method, this platform enables automated, cost-effective (0.66 USD per test), and high-performance testing of human cTnI in serum samples within 20 min with a detection range of 6 orders of magnitude, detection limit of 0.84 pg mL-1 (595-fold higher than conventional cTnI-LFA), and a coefficient of variation of 2.9-8.5%, which are comparable to the standard equipment and acceptable for clinical use. Moreover, cTnI analysis results using clinical serum/plasma samples revealed a strong correlation (R2 = 0.991) with contemporary standard equipment, demonstrating the practical application of this platform for high-performance POCT.
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