FARKA, Zdeněk, Matthias Jürgen MICKERT, Antonín HLAVÁČEK, Petr SKLÁDAL and Hans-Heiner GORRIS. Photon-upconversion nanoparticles for single-molecule immunoassays. In EUROPT(R)ODE 2021. 2021.
Other formats:   BibTeX LaTeX RIS
Basic information
Original name Photon-upconversion nanoparticles for single-molecule immunoassays
Authors FARKA, Zdeněk, Matthias Jürgen MICKERT, Antonín HLAVÁČEK, Petr SKLÁDAL and Hans-Heiner GORRIS.
Edition EUROPT(R)ODE 2021, 2021.
Other information
Original language English
Type of outcome Conference abstract
Field of Study 10406 Analytical chemistry
Country of publisher Poland
Confidentiality degree is not subject to a state or trade secret
Organization unit Faculty of Science
Tags International impact
Changed by Changed by: doc. Mgr. Zdeněk Farka, Ph.D., učo 357740. Changed: 4/12/2021 14:04.
Abstract
The recent progress in the field of immunoassays has been enabled by the development of various kinds of nanomaterials. In particular, photon-upconversion nanoparticles (UCNPs) proved to be excellent labels due to their ability to emit shorter wavelength light under near-infrared excitation (anti-Stokes emission), which prevents autofluorescence, minimizes light scattering, and thus reduces the optical background. These unique properties enabled imaging of single UCNPs by wide-field epiluminescence microscopy and allowed development of single-molecule (digital) immunoassays. We have synthesized conjugates of polyethylene glycol-coated UCNPs with streptavidin and applied them as a label for the detection of cancer biomarker prostate specific antigen (PSA). The detection was based on counting of individual sandwich immunocomplexes under the wide-field microscope. The noise-surpassing digital readout (particle counting) resulted in 20× higher sensitivity than the analog readout (output light intensity) and provided a detection limit of 23 fg mL−1 (800 aM) in 25% human serum. The conjugates of UCNPs with streptavidin are also useful as a label for the detection of bacterial pathogens. We have developed an immunoassay for the detection of causative agent of European foulbrood, bacterium Melissococcus plutonius. The assay provided a limit of detection of 340 CFU mL−1 and its practical applicability was verified by the analysis of real samples of bees, larvae and bottom hive debris.
Links
GA21-03156S, research and development projectName: Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů
Investor: Czech Science Foundation
LQ1601, research and development projectName: CEITEC 2020 (Acronym: CEITEC2020)
Investor: Ministry of Education, Youth and Sports of the CR
LTAB19011, research and development projectName: Luminiscenční imunostanovení jako citlivý nástroj pro diagnostiku nemocí včel
Investor: Ministry of Education, Youth and Sports of the CR, Sensitive bioanalytical tools for the surveillance of honey bee diseases, INTER-ACTION
PrintDisplayed: 31/8/2024 22:16