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@article{1394971,
author = {Farka, Zdeněk and Mickert, Matthias J. and Hlaváček, Antonín and Skládal, Petr and Gorris, Hans H.},
article_location = {Washington, DC},
article_number = {21},
doi = {http://dx.doi.org/10.1021/acs.analchem.7b03542},
keywords = {single-molecule analysis; upconversion-linked immunosorbent assay; cancer biomarker; prostate specific antigen; photon-upconversion nanoparticle},
language = {eng},
issn = {0003-2700},
journal = {Analytical Chemistry},
title = {Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers},
url = {http://pubs.acs.org/doi/10.1021/acs.analchem.7b03542},
volume = {89},
year = {2017}
}
TY - JOUR
ID - 1394971
AU - Farka, Zdeněk - Mickert, Matthias J. - Hlaváček, Antonín - Skládal, Petr - Gorris, Hans H.
PY - 2017
TI - Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers
JF - Analytical Chemistry
VL - 89
IS - 21
SP - 11825-11830
EP - 11825-11830
PB - American Chemical Society
SN - 00032700
KW - single-molecule analysis
KW - upconversion-linked immunosorbent assay
KW - cancer biomarker
KW - prostate specific antigen
KW - photon-upconversion nanoparticle
UR - http://pubs.acs.org/doi/10.1021/acs.analchem.7b03542
L2 - http://pubs.acs.org/doi/10.1021/acs.analchem.7b03542
N2 - The ability to detect disease markers at the single molecule level promises the ultimate sensitivity in clinical diagnosis. Fluorescence-based single-molecule analysis, however, is limited by matrix interference and can only probe a very small detection volume, which is typically not suitable for real world analytical applications. We have developed a microtiter plate immunoassay for counting single molecules of the cancer marker prostate specific antigen (PSA) using photon-upconversion nanoparticles (UCNPs) as labels that can be detected without background fluorescence. Individual sandwich immunocomplexes consisting of (1) an anti-PSA antibody immobilized to the surface of a microtiter well, (2) PSA, and (3) an anti-PSA antibody-UCNP conjugate were counted under a wide-field epifluorescence microscope equipped with a 980 nm laser excitation source. The single-molecule (digital) upconversion-linked immunosorbent assay (ULISA) reaches a limit of detection of 1.2 pg mL–1 (42 fM) PSA in 25% blood serum, which is about ten times more sensitive than commercial ELISAs, and covers a dynamic range of three orders of magnitude. This upconversion detection mode has the potential to pave the way for a new generation of digital immunoassays.
ER -
FARKA, Zdeněk, Matthias J. MICKERT, Antonín HLAVÁČEK, Petr SKLÁDAL and Hans H. GORRIS. Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers. \textit{Analytical Chemistry}. Washington, DC: American Chemical Society, 2017, vol.~89, No~21, p.~11825-11830. ISSN~0003-2700. Available from: https://dx.doi.org/10.1021/acs.analchem.7b03542.
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