Photon-upconversion nanoparticles as a background-free label in
rapid immunoassays for diclofenac

a 2018

Photon-upconversion nanoparticles as a background-free label in rapid immunoassays for diclofenac

FARKA, Zdeněk, Antonín HLAVÁČEK, Matthias Jürgen MICKERT, Dietmar KNOPP, Petr SKLÁDAL et. al.

Základní údaje

Originální název

Photon-upconversion nanoparticles as a background-free label in rapid immunoassays for diclofenac

Autoři

FARKA, Zdeněk, Antonín HLAVÁČEK, Matthias Jürgen MICKERT, Dietmar KNOPP, 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

photon-upconversion; upconversion-linked immunosorbent assay; bioconjugation; diclofenac

Příznaky

Mezinárodní význam

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

Anotace

V originále

Enzyme-linked immunosorbent assays (ELISAs) are widely used for detecting various analytes in complex matrices. However, they are limited by inherent instability of enzymes and time-consuming signal development. Thus, novel nanomaterial labels can offer several advantages, such as high stability and relatively easy preparation and storage. Photon-upconversion nanoparticles (UCNPs) are particularly suitable as background-free luminescent labels for immunoassays. We have developed an indirect and direct formats for the competitive upconversion-linked immunosorbent assay (ULISA). The target analyte was the pharmaceutical diclofenac (DCF), which is a common micropollutant in waters. The indirect assay involves a UCNP-labeled secondary antibody, while the tracer in the direct assay consists of UCNP enclosed by a carboxylated silica shell and coated with DCF-conjugated bovine γ-globulin. The protein coating prevents non-specific adsorption and provides a structurally flexible linker for surface-exposed DCF to allow efficient competition with free DCF in real water samples. The single-step direct assay has a limit of detection (LOD) of 20 pg mL−1 and reduces the analysis time to only 70 min. The optimized ULISA is comparable in sensitivity to the conventional ELISA while providing shorter analysis time of 70 min. Currently, we are able to visualize individual UCNP-labelled immunocomplexes, which provides essentially absolute sensitivity and allows to further enhance the immunoassay performance.

Návaznosti

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

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

Citovat

FARKA, Zdeněk, Antonín HLAVÁČEK, Matthias Jürgen MICKERT, Dietmar KNOPP, Petr SKLÁDAL a Hans-Heiner GORRIS. Photon-upconversion nanoparticles as a background-free label in rapid immunoassays for diclofenac. In UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials. 2018.

@proceedings{1414599,
   author = {Farka, Zdeněk and Hlaváček, Antonín and Mickert, Matthias Jürgen and Knopp, Dietmar and Skládal, Petr and Gorris, HansandHeiner},
   booktitle = {UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials},
   keywords = {photon-upconversion; upconversion-linked immunosorbent assay; bioconjugation; diclofenac},
   language = {eng},
   title = {Photon-upconversion nanoparticles as a background-free label in rapid immunoassays for diclofenac},
   year = {2018}
}

TY  - CONF
ID  - 1414599
AU  - Farka, Zdeněk - Hlaváček, Antonín - Mickert, Matthias Jürgen - Knopp, Dietmar - Skládal, Petr - Gorris, Hans-Heiner
PY  - 2018
TI  - Photon-upconversion nanoparticles as a background-free label in rapid immunoassays for diclofenac
KW  - photon-upconversion
KW  - upconversion-linked immunosorbent assay
KW  - bioconjugation
KW  - diclofenac
N2  - Enzyme-linked immunosorbent assays (ELISAs) are widely used for detecting various analytes in complex matrices. However, they are limited by inherent instability of enzymes and time-consuming signal development. Thus, novel nanomaterial labels can offer several advantages, such as high stability and relatively easy preparation and storage. Photon-upconversion nanoparticles (UCNPs) are particularly suitable as background-free luminescent labels for immunoassays. We have developed an indirect and direct formats for the competitive upconversion-linked immunosorbent assay (ULISA). The target analyte was the pharmaceutical diclofenac (DCF), which is a common micropollutant in waters. The indirect assay involves a UCNP-labeled secondary antibody, while the tracer in the direct assay consists of UCNP enclosed by a carboxylated silica shell and coated with DCF-conjugated bovine γ-globulin. The protein coating prevents non-specific adsorption and provides a structurally flexible linker for surface-exposed DCF to allow efficient competition with free DCF in real water samples. The single-step direct assay has a limit of detection (LOD) of 20 pg mL−1 and reduces the analysis time to only 70 min. The optimized ULISA is comparable in sensitivity to the conventional ELISA while providing shorter analysis time of 70 min. Currently, we are able to visualize individual UCNP-labelled immunocomplexes, which provides essentially absolute sensitivity and allows to further enhance the immunoassay performance.
ER  -

FARKA, Zdeněk, Antonín HLAVÁČEK, Matthias Jürgen MICKERT, Dietmar KNOPP, Petr SKLÁDAL a Hans-Heiner GORRIS. Photon-upconversion nanoparticles as a background-free label in rapid immunoassays for diclofenac. In \textit{UPCON 2018: 2nd Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials}. 2018.

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