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Original title:
Photon-upconversion nanoparticles for single-molecule immunosensing of cancer biomarkers and bacteria
Authors: Farka, Z. ; Mickert, M. J. ; Hlaváček, Antonín ; Poláchová, V. ; Kostiv, Uliana ; Pastucha, M. ; Horák, Daniel ; Gorris, H. H. ; Skládal, P.
Document type: Papers
Conference/Event: NANOCON 2019. International Conference on Nanomaterials - Research & Application /11./, Brno (CZ), 20191016
Year: 2020
Language: eng
Abstract: 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.
Keywords: bioconjugation; photon-upconversion nanoparticle; single-molecule detection
Project no.: LTAB19011
Funding provider: GA MŠk
Host item entry: NANOCON Conference Proceedings - International Conference on Nanomaterials, ISBN 978-808729495-6, ISSN 2694-930X
Note: Související webová stránka: https://www.confer.cz/nanocon/2019
Institution: Institute of Analytical Chemistry AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0311153
Permalink: http://www.nusl.cz/ntk/nusl-432155
The record appears in these collections:
Research > Institutes ASCR > Institute of Analytical Chemistry
Conference materials > Papers
Authors: Farka, Z. ; Mickert, M. J. ; Hlaváček, Antonín ; Poláchová, V. ; Kostiv, Uliana ; Pastucha, M. ; Horák, Daniel ; Gorris, H. H. ; Skládal, P.
Document type: Papers
Conference/Event: NANOCON 2019. International Conference on Nanomaterials - Research & Application /11./, Brno (CZ), 20191016
Year: 2020
Language: eng
Abstract: 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.
Keywords: bioconjugation; photon-upconversion nanoparticle; single-molecule detection
Project no.: LTAB19011
Funding provider: GA MŠk
Host item entry: NANOCON Conference Proceedings - International Conference on Nanomaterials, ISBN 978-808729495-6, ISSN 2694-930X
Note: Související webová stránka: https://www.confer.cz/nanocon/2019
Institution: Institute of Analytical Chemistry AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0311153
Permalink: http://www.nusl.cz/ntk/nusl-432155
The record appears in these collections:
Research > Institutes ASCR > Institute of Analytical Chemistry
Conference materials > Papers
Record created 2020-12-03, last modified 2023-12-06