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

a 2018

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

MICKERT, Matthias Jürgen, Zdeněk FARKA, Antonín HLAVÁČEK, Petr SKLÁDAL, Hans-Heiner GORRIS et. al.

Základní údaje

Originální název

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

Autoři

MICKERT, Matthias Jürgen, Zdeněk FARKA, Antonín HLAVÁČEK, Petr SKLÁDAL a Hans-Heiner GORRIS

Vydání

UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials, 2018

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10406 Analytical chemistry

Stát vydavatele

Španělsko

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

Single molecule; immunoassay; wide-field upconversion microscopy; cancer/disease biomarkers

Příznaky

Mezinárodní význam

Změněno: 26. 3. 2019 15:17, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

Detecting disease markers at the single molecule level has the potential to drastically improve the sensitivity in clinical diagnosis. Prostate specific antigen (PSA) is a reliable protein marker for prostate cancer, the most common form of cancer among men. After surgeries like radical prostatectomy PSA levels are very low. Detecting small increases in PSA concentration shortly after surgery helps to take the necessary steps to save the patient’s life. High photostability, near infrared excitation and biocompatibility after silanization make upconverting nanoparticles (UCNPs) a powerful tool for single molecule immunoassays. A conventional epifluorescence microscope (Nikon Eclipse Ti) was equipped with a 980 nm continuous wave laser diode (4 W) and a 100x objective with a NA of 1.49. Immunocomplexes consisting of (1) an anti-PSA capture antibody immobilized on the bottom of a microwell, (2) PSA, and (3) a silanized UCNP (NaYF4:Yb3+,Er3+) conjugated with a second anti-PSA antibody were counted under the microscope and additionally scanned in a microplate reader equipped with a 980 nm laser. The single molecule (digital) upconversion-linked immunosorbent assay (ULISA) reached a limit of detection of 1.2 pg ml-1 (42 fM) in 25% blood serum, which is about ten times more sensitive than the analog readout and commercial ELISAs. The dynamic range covered three orders of magnitude. The simple design and high sensitivity make this assay a pioneer for a new generation of digital immunoassays.

Návaznosti

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

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

Citovat

MICKERT, Matthias Jürgen, Zdeněk FARKA, Antonín HLAVÁČEK, Petr SKLÁDAL a Hans-Heiner GORRIS. Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers. In UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials. 2018.

@proceedings{1414600,
   author = {Mickert, Matthias Jürgen and Farka, Zdeněk and Hlaváček, Antonín and Skládal, Petr and Gorris, HansandHeiner},
   booktitle = {UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials},
   keywords = {Single molecule; immunoassay; wide-field upconversion microscopy; cancer/disease biomarkers},
   language = {eng},
   title = {Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers},
   year = {2018}
}

TY  - CONF
ID  - 1414600
AU  - Mickert, Matthias Jürgen - Farka, Zdeněk - Hlaváček, Antonín - Skládal, Petr - Gorris, Hans-Heiner
PY  - 2018
TI  - Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers
KW  - Single molecule
KW  - immunoassay
KW  - wide-field upconversion microscopy
KW  - cancer/disease biomarkers
N2  - Detecting disease markers at the single molecule level has the potential to drastically improve the sensitivity in clinical diagnosis. Prostate specific antigen (PSA) is a reliable protein marker for prostate cancer, the most common form of cancer among men. After surgeries like radical prostatectomy PSA levels are very low. Detecting small increases in PSA concentration shortly after surgery helps to take the necessary steps to save the patient’s life. High photostability, near infrared excitation and biocompatibility after silanization make upconverting nanoparticles (UCNPs) a powerful tool for single molecule immunoassays. A conventional epifluorescence microscope (Nikon Eclipse Ti) was equipped with a 980 nm continuous wave laser diode (4 W) and a 100x objective with a NA of 1.49. Immunocomplexes consisting of (1) an anti-PSA capture antibody immobilized on the bottom of a microwell, (2) PSA, and (3) a silanized UCNP (NaYF4:Yb3+,Er3+) conjugated with a second anti-PSA antibody were counted under the microscope and additionally scanned in a microplate reader equipped with a 980 nm laser. The single molecule (digital) upconversion-linked immunosorbent assay (ULISA) reached a limit of detection of 1.2 pg ml-1 (42 fM) in 25% blood serum, which is about ten times more sensitive than the analog readout and commercial ELISAs. The dynamic range covered three orders of magnitude. The simple design and high sensitivity make this assay a pioneer for a new generation of digital immunoassays.
ER  -

MICKERT, Matthias Jürgen, Zdeněk FARKA, Antonín HLAVÁČEK, Petr SKLÁDAL a Hans-Heiner GORRIS. Single molecule upconversion-linked immunosorbent assay with extended dynamic range for the sensitive detection of diagnostic biomarkers. In \textit{UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials}. 2018.

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