Farka, Zdenek and Mickert, Matthias J. and Hlavacek, Antonin and Skladal, Petr and Gorris, Hans H. (2017) Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers. ANALYTICAL CHEMISTRY, 89 (21). pp. 11825-11830. ISSN 0003-2700, 1520-6882
Full text not available from this repository. (Request a copy)Abstract
The ability to detect disease markers at the single molecule level promises the ultimate sensitivity in clinical diagnosis. Fluorescence-based single-molecule analysis, however, is limited by matrix interference and can only probe a very small detection volume, which is typically not suitable for real world analytical applications. We have developed a microtiter plate immunoassay for counting single molecules of the cancer marker prostate specific antigen (PSA) using photon-upconversion nanoparticles (UCNPs) as labels that can be detected without background fluorescence. Individual sandwich immunocomplexes consisting of (1) an anti-PSA antibody immobilized to the surface of a microtiter well, (2) PSA, and (3) an anti-PSA antibody-UCNP conjugate were counted under a wide-field epifluorescence microscope equipped with a 980 nm laser excitation source. The single-molecule (digital) upconversion-linked immunosorbent assay (ULISA) reaches a limit of detection of 1.2 pg mL(-1) (42 fM) PSA in 25% blood serum, which is about ten times more sensitive than commercial ELISAs, and covers a dynamic range of three orders of magnitude. This upconversion detection mode has the potential to pave the way for a new generation of digital immunoassays.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PHOTON-UPCONVERTING NANOPARTICLES; PROSTATE-SPECIFIC ANTIGEN; BIOANALYTICAL APPLICATIONS; EUROPIUM NANOPARTICLES; QUANTITATIVE-ANALYSIS; IMMUNOASSAY; CHALLENGES; CANCER; PERSPECTIVES; PHASE; |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 13:19 |
| Last Modified: | 20 Feb 2019 08:17 |
| URI: | https://pred.uni-regensburg.de/id/eprint/1859 |
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