Single Molecule Upconversion-Linked Immunosorbent Assay with
Extended Dynamic Range for the Sensitive Detection of
Diagnostic Biomarkers

J 2017

Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers

FARKA, Zdeněk, Matthias J. MICKERT, Antonín HLAVÁČEK, Petr SKLÁDAL, Hans H. GORRIS et. al.

Základní údaje

Originální název

Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers

Autoři

FARKA, Zdeněk (203 Česká republika, domácí), Matthias J. MICKERT (276 Německo), Antonín HLAVÁČEK (203 Česká republika, domácí), Petr SKLÁDAL (203 Česká republika, domácí) a Hans H. GORRIS (276 Německo, garant)

Vydání

Analytical Chemistry, Washington, DC, American Chemical Society, 2017, 0003-2700

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 6.042

Kód RIV

RIV/00216224:14740/17:00095148

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1021/acs.analchem.7b03542

UT WoS

000414887000093

Klíčová slova anglicky

single-molecule analysis; upconversion-linked immunosorbent assay; cancer biomarker; prostate specific antigen; photon-upconversion nanoparticle

Štítky

CF CRYO, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 15. 3. 2018 12:44, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

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.

Návaznosti

GBP206/12/G014, projekt VaV

Název: Centrum pokročilých bioanalytických technologií

LD15023, projekt VaV

Název: Příprava diskrétních biokonjugátů photon-upconversion nanočástic

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Příprava diskrétních biokonjugátů photon-upconversion nanočástic

LM2015043, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

Citovat

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. 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.

@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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