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Urchin peroxidase-mimicking Au@Pt nanoparticles as a label in lateral flow immunoassay: impact of nanoparticle composition on detection limit of Clavibacter michiganensis

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Abstract

The influence of Au@Pt nanoparticles’ composition, morphology, and peroxidase-mimicking activity on the limit of detection (LOD) of lateral flow immunoassay (LFIA) has been investigated. Fourteen types of nanoparticles were synthesized by varying the concentration of Pt4+ (20–2000 μM), using gold nanoparticles (GNP, diameter 20.0 ± 2.6 nm) as the seeds and ascorbic acid as a reducing agent. Au@Pt nanoparticles and GNPs were conjugated with antibodies specific to the target analyte, a widespread and dangerous phytopathogenic bacteria species (Clavibacter michiganensis). We found that the 100-fold growth of the Pt4+ concentration was accompanied by an increase of the Au@Pt nanoparticle diameter (24–55 nm) and surface area with the formation of urchin-shaped morphology. These changes led to a 70-fold increase in peroxidase-mimicking activity in the solution (specific activity 0.06–4.4 U mg−1) and a 30-fold decrease in LOD using the catalytic activity of Au@Pt. The Au@Pt nanoparticles synthesized at 1000–2000 μM of Pt4+ demonstrated statistically indistinguishable catalytic activity. The highest sensitivity of LFIA was reached for Au@Pt nanoparticles synthesized at Pt4+ concentration equal to 1000 μM. Au@Pt nanoparticles saved most of their peroxidase-mimicking activity, whereas endogenous plant peroxidases were completely inhibited by sodium azide. The LOD of LFIA with Au@Pt nanoparticles synthesized at 1200 μM of Pt4+ was 300 colony-forming units (CFU) per mL of buffer and 500 CFU per mL of potato tuber extract, which provides 330- and 200-fold improvement compared to the conventional LFIA with GNPs. The assay consists of three rapid 5-min stages, namely, extraction, lateral flow, and color enhancement (oxidation of diaminobenzidine by Au@Pt nanoparticles). LFIA with the urchin Au@Pt nanoparticles allows the detection of latent bacterial infections rapidly without equipment or special skills.

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Acknowledgments

The authors are grateful to Yu.A. Varitsev (A.G. Lorch All-Russian Potato Research Institute, Korenevo, Moscow region, Russia) for providing the infected and healthy plants, N.V. Drenova (All-Russian Plant Quarantine Centre, Bykovo-2, Moscow region, Russia) for providing bacterial strains, and E.S. Slutskaya (Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences, Moscow, Russia) for the ICP MS measurements.

Funding

This study was financially supported by the Russian Science Foundation (grant no. 16-16-04108).

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Authors and Affiliations

  1. A.N. Bach Institute of Biochemistry, Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences, Leninsky prospect 33, 119071, Moscow, Russia

    Vasily G. Panferov, Irina V. Safenkova, Anatoly V. Zherdev & Boris B. Dzantiev

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  1. Vasily G. Panferov
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  2. Irina V. Safenkova
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  3. Anatoly V. Zherdev
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Correspondence to Boris B. Dzantiev.

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Panferov, V.G., Safenkova, I.V., Zherdev, A.V. et al. Urchin peroxidase-mimicking Au@Pt nanoparticles as a label in lateral flow immunoassay: impact of nanoparticle composition on detection limit of Clavibacter michiganensis. Microchim Acta 187, 268 (2020). https://doi.org/10.1007/s00604-020-04253-3

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