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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 a 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, roč.~89, č.~21, s.~11825-11830. ISSN~0003-2700. Dostupné z: https://dx.doi.org/10.1021/acs.analchem.7b03542.
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