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. 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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Basic information
Original name Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers
Authors FARKA, Zdeněk (203 Czech Republic, belonging to the institution), Matthias J. MICKERT (276 Germany), Antonín HLAVÁČEK (203 Czech Republic, belonging to the institution), Petr SKLÁDAL (203 Czech Republic, belonging to the institution) and Hans H. GORRIS (276 Germany, guarantor).
Edition Analytical Chemistry, Washington, DC, American Chemical Society, 2017, 0003-2700.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10406 Analytical chemistry
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 6.042
RIV identification code RIV/00216224:14740/17:00095148
Organization unit Central European Institute of Technology
Doi http://dx.doi.org/10.1021/acs.analchem.7b03542
UT WoS 000414887000093
Keywords in English single-molecule analysis; upconversion-linked immunosorbent assay; cancer biomarker; prostate specific antigen; photon-upconversion nanoparticle
Tags CF CRYO, rivok
Tags International impact, Reviewed
Changed by Changed by: Mgr. Pavla Foltynová, Ph.D., učo 106624. Changed: 15/3/2018 12:44.
Abstract
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.
Links
GBP206/12/G014, research and development projectName: Centrum pokročilých bioanalytických technologií
LD15023, research and development projectName: Příprava diskrétních biokonjugátů photon-upconversion nanočástic
Investor: Ministry of Education, Youth and Sports of the CR
LM2015043, research and development projectName: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
LQ1601, research and development projectName: CEITEC 2020 (Acronym: CEITEC2020)
Investor: Ministry of Education, Youth and Sports of the CR
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