Název:
Photon-upconversion nanoparticles for single-molecule immunosensing of cancer biomarkers and bacteria
Autoři:
Farka, Z. ; Mickert, M. J. ; Hlaváček, Antonín ; Poláchová, V. ; Kostiv, Uliana ; Pastucha, M. ; Horák, Daniel ; Gorris, H. H. ; Skládal, P. Typ dokumentu: Příspěvky z konference Konference/Akce: NANOCON 2019. International Conference on Nanomaterials - Research & Application /11./, Brno (CZ), 20191016
Rok:
2020
Jazyk:
eng
Abstrakt: The recent progress in the field of immunoassays has been driven by introduction of various kinds of nanomaterials. In particular, photon-upconversion nanoparticles (UCNPs) proved to be excellent immunoassay labels due to their ability to emit light of shorter wavelengths under near-infrared excitation (anti-Stokes emission), which prevents autofluorescence, minimizes light scattering, and thus reduces the optical background interference. These unique photoluminescent properties allow counting of individual biomolecules labeled with UCNPs by conventional wide-field epiluminescence microscopy and enable the development of single-molecule (digital) immunoassays. We have introduced a novel label based on UCNPs conjugated with streptavidin via poly(ethylene glycol) and applied it in a digital upconversion-linked immunosorbent assay (ULISA) for the detection of a cancer biomarker prostate specific antigen (PSA). The digital readout based on counting of individual immunocomplexes improved the sensitivity 16× compared to conventional analog readout and allowed to reach a limit of detection (LOD) of 23 fg·mL−1 (800 aM). Human serum samples were successfully analyzed achieving an excellent correlation with electrochemiluminescence reference method. The conjugates of UCNPs with streptavidin are also suitable for the detection of pathogenic bacterium Melissococcus plutonius, the causative agent of honeybee disease European foulbrood. The ULISA assay provided an LOD of 340 CFU·mL−1 and successfully analyzed real samples of bees, larvae and bottom hive debris. Due to the high reliability and relatively simple detection scheme, the digital ULISA can pave the way for a new generation of digital immunoassays with a strong potential for commercialization.
Klíčová slova:
bioconjugation; photon-upconversion nanoparticle; single-molecule detection Číslo projektu: LTAB19011 Poskytovatel projektu: GA MŠk Zdrojový dokument: NANOCON Conference Proceedings - International Conference on Nanomaterials, ISBN 978-808729495-6, ISSN 2694-930X Poznámka: Související webová stránka: https://www.confer.cz/nanocon/2019
Instituce: Ústav analytické chemie AV ČR
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Informace o dostupnosti dokumentu:
Dokument je dostupný na vyžádání prostřednictvím repozitáře Akademie věd. Původní záznam: http://hdl.handle.net/11104/0311153