Digital detection of subfemtomolar concentrations of
prostate-specific antigen by single molecule immunosensing

a 2021

Digital detection of subfemtomolar concentrations of prostate-specific antigen by single molecule immunosensing

GORRIS, Hans-Heiner, Matthias Jürgen MICKERT, Zdeněk FARKA, Uliana KOSTIV, Antonín HLAVÁČEK et. al.

Základní údaje

Originální název

Digital detection of subfemtomolar concentrations of prostate-specific antigen by single molecule immunosensing

Autoři

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

Vydání

31st Anniversary World Congress on Biosensors, 2021

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10406 Analytical chemistry

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Přírodovědecká fakulta

Příznaky

Mezinárodní význam

Změněno: 2. 8. 2021 16:04, doc. Mgr. Zdeněk Farka, Ph.D.

Anotace

V originále

The ability to detect single molecules holds the promise to reach the ultimate sensitivity in analytical chemistry. Several problems, however, must be addressed before single-molecule techniques can be employed for routine analytical applications. Photon-upconversion nanoparticles (UCNPs) are excellent labels for single molecule immunoassays because they emit short-wavelength light under near-infrared (NIR, 980 nm) excitation (anti-Stokes emission), which avoids autofluorescence and light scattering. These unique photoluminescent features enable the detection of UCNPs at the single nanoparticle level by conventional wide-field epiluminescence microscopy. Furthermore, very homogeneous UCNP labels were prepared by gel electrophoretic separation. For the upconversion-linked immunoassay (ULISA), microtiter plates were first coated with anti-prostate-specific antigen (PSA) antibodies to capture PSA. PSA was then detected either by an anti-PSA antibody-UCNP conjugate or a biotinylated anti-PSA antibody in combination with a streptavidin-PEG UCNP conjugate. The PSA concentration was determined on the same microtiter plate either (1) by using an upconversion microtiter plate reader (analog mode) or (2) by counting individual immunocomplexes under an upconversion wide-field microscope (digital mode). The lowest limit of detection was achieved by using the streptavidin-PEG-neridronate conjugate as a detection label. Counting single analyte molecules improved the limit of detection (LOD) (23 fg mL-1, 800 aM) by more than one order of magnitude compared to the analogue readout.

Návaznosti

GA21-03156S, interní kód MU

Název: Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů

Investor: Grantová agentura ČR, Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

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

LTAB19011, projekt VaV

Název: Luminiscenční imunostanovení jako citlivý nástroj pro diagnostiku nemocí včel

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Luminiscenční imunostanovení jako citlivý nástroj pro diagnostiku nemocí včel, INTER-ACTION

Citovat

GORRIS, Hans-Heiner, Matthias Jürgen MICKERT, Zdeněk FARKA, Uliana KOSTIV, Antonín HLAVÁČEK, Daniel HORÁK a Petr SKLÁDAL. Digital detection of subfemtomolar concentrations of prostate-specific antigen by single molecule immunosensing. In 31st Anniversary World Congress on Biosensors. 2021.

@proceedings{1784817,
   author = {Gorris, HansandHeiner and Mickert, Matthias Jürgen and Farka, Zdeněk and Kostiv, Uliana and Hlaváček, Antonín and Horák, Daniel and Skládal, Petr},
   booktitle = {31st Anniversary World Congress on Biosensors},
   language = {eng},
   title = {Digital detection of subfemtomolar concentrations of prostate-specific antigen by single molecule immunosensing},
   year = {2021}
}

TY  - CONF
ID  - 1784817
AU  - Gorris, Hans-Heiner - Mickert, Matthias Jürgen - Farka, Zdeněk - Kostiv, Uliana - Hlaváček, Antonín - Horák, Daniel - Skládal, Petr
PY  - 2021
TI  - Digital detection of subfemtomolar concentrations of prostate-specific antigen by single molecule immunosensing
N2  - The ability to detect single molecules holds the promise to reach the ultimate sensitivity in analytical chemistry. Several problems, however, must be addressed before single-molecule techniques can be employed for routine analytical applications. Photon-upconversion nanoparticles (UCNPs) are excellent labels for single molecule immunoassays because they emit short-wavelength light under near-infrared (NIR, 980 nm) excitation (anti-Stokes emission), which avoids autofluorescence and light scattering. These unique photoluminescent features enable the detection of UCNPs at the single nanoparticle level by conventional wide-field epiluminescence microscopy. Furthermore, very homogeneous UCNP labels were prepared by gel electrophoretic separation. For the upconversion-linked immunoassay (ULISA), microtiter plates were first coated with anti-prostate-specific antigen (PSA) antibodies to capture PSA. PSA was then detected either by an anti-PSA antibody-UCNP conjugate or a biotinylated anti-PSA antibody in combination with a streptavidin-PEG UCNP conjugate. The PSA concentration was determined on the same microtiter plate either (1) by using an upconversion microtiter plate reader (analog mode) or (2) by counting individual immunocomplexes under an upconversion wide-field microscope (digital mode). The lowest limit of detection was achieved by using the streptavidin-PEG-neridronate conjugate as a detection label. Counting single analyte molecules improved the limit of detection (LOD) (23 fg mL-1, 800 aM) by more than one order of magnitude compared to the analogue readout.
ER  -

GORRIS, Hans-Heiner, Matthias Jürgen MICKERT, Zdeněk FARKA, Uliana KOSTIV, Antonín HLAVÁČEK, Daniel HORÁK a Petr SKLÁDAL. Digital detection of subfemtomolar concentrations of prostate-specific antigen by single molecule immunosensing. In \textit{31st Anniversary World Congress on Biosensors}. 2021.

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