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

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

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

EID Scopus

2-s2.0-85043459122

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

Cite

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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