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.

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

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

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

Abstract

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.

Links

GBP206/12/G014, research and development project

Name: Centrum pokročilých bioanalytických technologií

LD15023, research and development project

Name: Příprava diskrétních biokonjugátů photon-upconversion nanočástic

Investor: Ministry of Education, Youth and Sports of the CR

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

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.

@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 and 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, 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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