Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081715%3A_____%2F17%3A00481051" target="_blank" >RIV/68081715:_____/17:00481051 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14740/17:00095148

Výsledek na webu

<a href="http://dx.doi.org/10.1021/acs.analchem.7b03542" target="_blank" >http://dx.doi.org/10.1021/acs.analchem.7b03542</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acs.analchem.7b03542" target="_blank" >10.1021/acs.analchem.7b03542</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers

Popis výsledku v původním jazyce

The ability to detect disease markers at the single molecule level promisesnthe ultimate sensitivity in clinical diagnosis. Fluorescence-based single-molecule analysis,nhowever, is limited by matrix interference and can only probe a very small detectionnvolume, which is typically not suitable for real world analytical applications. We havendeveloped a microtiter plate immunoassay for counting single molecules of the cancernmarker prostate specific antigen (PSA) using photon-upconversion nanoparticlesn(UCNPs) as labels that can be detected without background fluorescence. Individualnsandwich immunocomplexes consisting of (1) an anti-PSA antibody immobilized to thensurface of a microtiter well, (2) PSA, and (3) an anti-PSA antibody-UCNP conjugate werencounted under a wide-field epifluorescence microscope equipped with a 980 nm lasernexcitation source. The single-molecule (digital) upconversion-linked immunosorbent assayn(ULISA) reaches a limit of detection of 1.2 pg mL−1 (42 fM) PSA in 25% blood serum,nwhich is about ten times more sensitive than commercial ELISAs, and covers a dynamicnrange of three orders of magnitude. This upconversion detection mode has the potentialnto pave the way for a new generation of digital immunoassays.

Název v anglickém jazyce

Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers

Popis výsledku anglicky

The ability to detect disease markers at the single molecule level promisesnthe ultimate sensitivity in clinical diagnosis. Fluorescence-based single-molecule analysis,nhowever, is limited by matrix interference and can only probe a very small detectionnvolume, which is typically not suitable for real world analytical applications. We havendeveloped a microtiter plate immunoassay for counting single molecules of the cancernmarker prostate specific antigen (PSA) using photon-upconversion nanoparticlesn(UCNPs) as labels that can be detected without background fluorescence. Individualnsandwich immunocomplexes consisting of (1) an anti-PSA antibody immobilized to thensurface of a microtiter well, (2) PSA, and (3) an anti-PSA antibody-UCNP conjugate werencounted under a wide-field epifluorescence microscope equipped with a 980 nm lasernexcitation source. The single-molecule (digital) upconversion-linked immunosorbent assayn(ULISA) reaches a limit of detection of 1.2 pg mL−1 (42 fM) PSA in 25% blood serum,nwhich is about ten times more sensitive than commercial ELISAs, and covers a dynamicnrange of three orders of magnitude. This upconversion detection mode has the potentialnto pave the way for a new generation of digital immunoassays.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

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OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Analytical Chemistry

ISSN

0003-2700

e-ISSN

—

Svazek periodika

89

Číslo periodika v rámci svazku

NOV

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

6

Strana od-do

11825-11830

Kód UT WoS článku

000414887000093

EID výsledku v databázi Scopus

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