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@proceedings{1807139,
author = {Farka, Zdeněk and Mickert, Matthias Jürgen and Hlaváček, Antonín and Skládal, Petr and Gorris, HansandHeiner},
booktitle = {EUROPT(R)ODE 2021},
language = {eng},
title = {Photon-upconversion nanoparticles for single-molecule immunoassays},
year = {2021}
}
TY - CONF
ID - 1807139
AU - Farka, Zdeněk - Mickert, Matthias Jürgen - Hlaváček, Antonín - Skládal, Petr - Gorris, Hans-Heiner
PY - 2021
TI - Photon-upconversion nanoparticles for single-molecule immunoassays
N2 - The recent progress in the field of immunoassays has been enabled by the development of various kinds of nanomaterials. In particular, photon-upconversion nanoparticles (UCNPs) proved to be excellent labels due to their ability to emit shorter wavelength light under near-infrared excitation (anti-Stokes emission), which prevents autofluorescence, minimizes light scattering, and thus reduces the optical background. These unique properties enabled imaging of single UCNPs by wide-field epiluminescence microscopy and allowed development of single-molecule (digital) immunoassays. We have synthesized conjugates of polyethylene glycol-coated UCNPs with streptavidin and applied them as a label for the detection of cancer biomarker prostate specific antigen (PSA). The detection was based on counting of individual sandwich immunocomplexes under the wide-field microscope. The noise-surpassing digital readout (particle counting) resulted in 20× higher sensitivity than the analog readout (output light intensity) and provided a detection limit of 23 fg mL−1 (800 aM) in 25% human serum. The conjugates of UCNPs with streptavidin are also useful as a label for the detection of bacterial pathogens. We have developed an immunoassay for the detection of causative agent of European foulbrood, bacterium Melissococcus plutonius. The assay provided a limit of detection of 340 CFU mL−1 and its practical applicability was verified by the analysis of real samples of bees, larvae and bottom hive debris.
ER -
FARKA, Zdeněk, Matthias Jürgen MICKERT, Antonín HLAVÁČEK, Petr SKLÁDAL a Hans-Heiner GORRIS. Photon-upconversion nanoparticles for single-molecule immunoassays. In \textit{EUROPT(R)ODE 2021}. 2021.
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