Self-Assembling Allochroic Nanocatalyst for Improving Nanozyme-Based Immunochromatographic Assays

Abstract

Paper-based rapid diagnostic tests, such as immunochromatographic assays, namely lateral flow immunoassay (LFA), are valuable alternatives for biomarker detection compared to traditional laboratory-based tests, but these assays need further refinement to consolidate their biosensing capabilities. Nanozyme integration into LFA systems may provide a reliable means of improving the analytic sensitivity of LFA tests. Due to the involvement of multiple liquid-handling steps, the quantitative accuracy is compromised, hence hindering the use of untrained personnel point-of-care use. Self-assembling allochroic nanocatalyst (SAN) assemblies satisfy these LFA quality measures by optimizing analyte-antibody reporting performance and by intrinsically catalyzing chromogen activation, thereby reducing the number of liquid handling steps involved during sample analysis. In SANs, the hydrophobic chromogens serve as peroxidase substrates that self-assemble into nanoparticles at high loading fractions. These features demonstrate the potential for SAN-LFAs to be a valuable patient point-of-care (POC) test. Herein, we describe the SAN fabrication process and employ SAN-LFAs to detect cardiac troponin I-troponin C (cTnI-TnC) and myoglobin (Myo) levels present in plasma samples. Using SAN-LFAs, the limits of detection for cTnI-TnC and Myo were 0.012 ng/mL and 0.2 ng/mL respectively. We also demonstrate SAN compatibility with blood samples and stability under long-term storage conditions. The successful utlization of SANs in LFA-based biomarker detection may inspire these nanocatalysts to be integrated into similar immunochromatographic testing methods.