Measurement of Sub-femtomolar Concentrations of
Prostate-Specific Antigen through Single-Molecule Counting with
an Upconversion-Linked Immunosorbent Assay

J 2019

Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay

MICKERT, Matthias Jürgen; Zdeněk FARKA; Uliana KOSTIV; Antonín HLAVÁČEK; Daniel HORÁK et al.

Základní údaje

Originální název

Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay

Autoři

MICKERT, Matthias Jürgen; Zdeněk FARKA; Uliana KOSTIV; Antonín HLAVÁČEK; Daniel HORÁK; Petr SKLÁDAL a Hans-Heiner GORRIS

Vydání

Analytical Chemistry, Washington, DC, American Chemical Society, 2019, 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: 6.785

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/19:00110348

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

photon-upconversion nanoparticle; bioconjugation; single-molecule immunoassay; digital detection; prostate-specific antigen; cancer biomarker

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 24. 10. 2024 12:29, Mgr. Adéla Pešková

Anotace

V originále

Single-molecule (digital) immunoassays provide the ability to detect much lower protein concentrations than conventional immunoassays. As photon-upconversion nanoparticles (UCNPs) can be detected without optical background interference, they are excellent labels for so-called single-molecule upconversion-linked immunosorbent assays (ULISAs). We have introduced a UCNP label design based on streptavidin-PEG-neridronate and a two-step detection scheme involving a biotinylated antibody that efficiently reduces nonspecific binding on microtiter plates. In a microtiter plate immunoassay, individual sandwich immune complexes of the cancer marker prostate-specific antigen (PSA) are detected and counted by wide-field epiluminescence microscopy (digital readout). The digital detection is 16× more sensitive than the respective analogue readout and thus expands the limit of detection to the sub-femtomolar concentration range (LOD: 23 fg mL–1, 800 aM). The single molecule ULISA shows excellent correlation with an electrochemiluminescence reference method. Although the analogue readout can routinely measure PSA concentrations in human serum samples, very low concentrations have to be monitored after radical prostatectomy. Combining the digital and analogue readout covers a dynamic range of more than 3 orders of magnitude in a single experiment.

Návaznosti

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

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

90043, velká výzkumná infrastruktura

Název: CIISB

Citovat

MICKERT, Matthias Jürgen; Zdeněk FARKA; Uliana KOSTIV; Antonín HLAVÁČEK; Daniel HORÁK; Petr SKLÁDAL a Hans-Heiner GORRIS. Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay. Analytical Chemistry. Washington, DC: American Chemical Society, 2019, roč. 91, č. 15, s. 9435-9441. ISSN 0003-2700. Dostupné z: https://doi.org/10.1021/acs.analchem.9b02872.

@article{1548841,
   author = {Mickert, Matthias Jürgen and Farka, Zdeněk and Kostiv, Uliana and Hlaváček, Antonín and Horák, Daniel and Skládal, Petr and Gorris, HansandHeiner},
   article_location = {Washington, DC},
   article_number = {15},
   doi = {https://doi.org/10.1021/acs.analchem.9b02872},
   keywords = {photon-upconversion nanoparticle; bioconjugation; single-molecule immunoassay; digital detection; prostate-specific antigen; cancer biomarker},
   language = {eng},
   issn = {0003-2700},
   journal = {Analytical Chemistry},
   title = {Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay},
   url = {https://pubs.acs.org/doi/10.1021/acs.analchem.9b02872},
   volume = {91},
   year = {2019}
}

TY  - JOUR
ID  - 1548841
AU  - Mickert, Matthias Jürgen - Farka, Zdeněk - Kostiv, Uliana - Hlaváček, Antonín - Horák, Daniel - Skládal, Petr - Gorris, Hans-Heiner
PY  - 2019
TI  - Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay
JF  - Analytical Chemistry
VL  - 91
IS  - 15
SP  - 9435-9441
EP  - 9435-9441
PB  - American Chemical Society
SN  - 00032700
KW  - photon-upconversion nanoparticle
KW  - bioconjugation
KW  - single-molecule immunoassay
KW  - digital detection
KW  - prostate-specific antigen
KW  - cancer biomarker
UR  - https://pubs.acs.org/doi/10.1021/acs.analchem.9b02872
L2  - https://pubs.acs.org/doi/10.1021/acs.analchem.9b02872
N2  - Single-molecule (digital) immunoassays provide the ability to detect much lower protein concentrations than conventional immunoassays. As photon-upconversion nanoparticles (UCNPs) can be detected without optical background interference, they are excellent labels for so-called single-molecule upconversion-linked immunosorbent assays (ULISAs). We have introduced a UCNP label design based on streptavidin-PEG-neridronate and a two-step detection scheme involving a biotinylated antibody that efficiently reduces nonspecific binding on microtiter plates. In a microtiter plate immunoassay, individual sandwich immune complexes of the cancer marker prostate-specific antigen (PSA) are detected and counted by wide-field epiluminescence microscopy (digital readout). The digital detection is 16× more sensitive than the respective analogue readout and thus expands the limit of detection to the sub-femtomolar concentration range (LOD: 23 fg mL–1, 800 aM). The single molecule ULISA shows excellent correlation with an electrochemiluminescence reference method. Although the analogue readout can routinely measure PSA concentrations in human serum samples, very low concentrations have to be monitored after radical prostatectomy. Combining the digital and analogue readout covers a dynamic range of more than 3 orders of magnitude in a single experiment.
ER  -

MICKERT, Matthias Jürgen; Zdeněk FARKA; Uliana KOSTIV; Antonín HLAVÁČEK; Daniel HORÁK; Petr SKLÁDAL a Hans-Heiner GORRIS. Measurement of Sub-femtomolar Concentrations of Prostate-Specific Antigen through Single-Molecule Counting with an Upconversion-Linked Immunosorbent Assay. \textit{Analytical Chemistry}. Washington, DC: American Chemical Society, 2019, roč.~91, č.~15, s.~9435-9441. ISSN~0003-2700. Dostupné z: https://doi.org/10.1021/acs.analchem.9b02872.

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