| MICKERT, Matthias Jürgen, Zdeněk FARKA, Uliana KOSTIV, Antonín HLAVÁČEK, Daniel HORÁK, Petr SKLÁDAL a Hans-Heiner GORRIS. Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay. Analytical Chemistry. Washington, DC: American Chemical Society, 2019, roč. 91, č. 15, s. 9435-9441. ISSN 0003-2700. Dostupné z: https://dx.doi.org/10.1021/acs.analchem.9b02872. |
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@article{1548841,
author = {Mickert, Matthias Jürgen and Farka, Zdeněk and Kostiv, Uliana and Hlaváček, Antonín and Horák, Daniel and Skládal, Petr and Gorris, HansandHeiner},
article_location = {Washington, DC},
article_number = {15},
doi = {http://dx.doi.org/10.1021/acs.analchem.9b02872},
keywords = {photon-upconversion nanoparticle; bioconjugation; single-molecule immunoassay; digital detection; prostate-specific antigen; cancer biomarker},
language = {eng},
issn = {0003-2700},
journal = {Analytical Chemistry},
title = {Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay},
url = {https://pubs.acs.org/doi/10.1021/acs.analchem.9b02872},
volume = {91},
year = {2019}
}
TY - JOUR
ID - 1548841
AU - Mickert, Matthias Jürgen - Farka, Zdeněk - Kostiv, Uliana - Hlaváček, Antonín - Horák, Daniel - Skládal, Petr - Gorris, Hans-Heiner
PY - 2019
TI - Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay
JF - Analytical Chemistry
VL - 91
IS - 15
SP - 9435-9441
EP - 9435-9441
PB - American Chemical Society
SN - 00032700
KW - photon-upconversion nanoparticle
KW - bioconjugation
KW - single-molecule immunoassay
KW - digital detection
KW - prostate-specific antigen
KW - cancer biomarker
UR - https://pubs.acs.org/doi/10.1021/acs.analchem.9b02872
L2 - https://pubs.acs.org/doi/10.1021/acs.analchem.9b02872
N2 - Single-molecule (digital) immunoassays provide the ability to detect much lower protein concentrations than conventional immunoassays. As photon-upconversion nanoparticles (UCNPs) can be detected without optical background interference, they are excellent labels for so-called single-molecule upconversion-linked immunosorbent assays (ULISAs). We have introduced a UCNP label design based on streptavidin-PEG-neridronate and a two-step detection scheme involving a biotinylated antibody that efficiently reduces nonspecific binding on microtiter plates. In a microtiter plate immunoassay, individual sandwich immune complexes of the cancer marker prostate-specific antigen (PSA) are detected and counted by wide-field epiluminescence microscopy (digital readout). The digital detection is 16× more sensitive than the respective analogue readout and thus expands the limit of detection to the sub-femtomolar concentration range (LOD: 23 fg mL–1, 800 aM). The single molecule ULISA shows excellent correlation with an electrochemiluminescence reference method. Although the analogue readout can routinely measure PSA concentrations in human serum samples, very low concentrations have to be monitored after radical prostatectomy. Combining the digital and analogue readout covers a dynamic range of more than 3 orders of magnitude in a single experiment.
ER -
MICKERT, Matthias Jürgen, Zdeněk FARKA, Uliana KOSTIV, Antonín HLAVÁČEK, Daniel HORÁK, Petr SKLÁDAL a Hans-Heiner GORRIS. Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay. \textit{Analytical Chemistry}. Washington, DC: American Chemical Society, 2019, roč.~91, č.~15, s.~9435-9441. ISSN~0003-2700. Dostupné z: https://dx.doi.org/10.1021/acs.analchem.9b02872.
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