The steadfast Au@Pt soldier: Peroxide-tolerant nanozyme for signal enhancement in lateral flow immunoassay of peroxidase-containing samples

Abstract

We report the approach for the detection of Au@Pt core@shell nanoparticles (nanozymes) with peroxidase-mimicking activity (PMA) in samples with high endogenous peroxidase activity (EPA). Unlike the endogenous peroxidases in plant extracts that are inhibited by elevated H2O2 (>20 mM), the PMA of nanozymes was stable in concentrated H2O2 (up to 4 M). Such a different stability of enzymes and Au@Pt to the substrate allowed for eliminating EPA and detecting only nanozymes. The developed approach was used for reaching a lower limit of detection (LOD) and eliminating the background for the lateral flow immunoassay (LFIA) of the important plant pathogen potato virus X (PVX) in leaf and tuber extracts. Using the PMA of Au@Pt, the LOD was reduced to 4 and 8 pg/mL in tuber and leaf extracts, respectively. The LOD values are 250- and 500-times lower in comparison with LFIA with conventional gold nanoparticles. The developed approach of peroxidases inhibition is universal for bioanalytical methods, and its applicability was confirmed by the elimination of EPA in three matrixes (serum, potato leaf and tuber extracts).