Upconversion Fluorescence Enables Homogeneous Immunoassay in Whole Blood
| Autor: | Timo Lövgren, Tero Soukka, Telle Ukonaho, Katri Kuningas, Terhi Rantanen, Henna Päkkilä |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Analyte
Clinical Biochemistry Analytical chemistry Phosphor 02 engineering and technology 010402 general chemistry Photochemistry Lanthanoid Series Elements 01 natural sciences 7. Clean energy Fluorescence Fluorescence Resonance Energy Transfer medicine Humans Immunoassay Luminescent Agents Estradiol medicine.diagnostic_test Chemistry Biochemistry (medical) 021001 nanoscience & nanotechnology Acceptor Photon upconversion 0104 chemical sciences Autofluorescence Förster resonance energy transfer 0210 nano-technology Blood Chemical Analysis |
| Zdroj: | Clinical Chemistry. 53:145-146 |
| ISSN: | 1530-8561 0009-9147 |
| Popis: | Many medical conditions are related to alterations in protein or hormone concentrations in blood. These changes are commonly detected with heterogeneous immunoassays that require a separation step before signal measurement. A simple homogeneous assay performable directly in whole blood would be useful in clinical diagnostics, but the performance of homogeneous fluorescence-based immunoassays has been severely limited by autofluorescence and strong absorption of ultraviolet and visible light in whole blood. These problems might be avoided by the use of near-infrared excited upconversion fluorescence. We recently described a novel homogeneous assay method based on upconversion fluorescence resonance energy transfer (FRET). In this method an upconverting phosphor (UCP) is used as a donor and a fluorescent protein or a small-molecular fluorescent dye is used as an acceptor (1)(2). We report here the use of a competitive homogeneous immunoassay for 17β-estradiol (E2) in whole blood as a model to demonstrate the application of upconversion FRET using a near-infrared fluorescent acceptor dye (Fig. 1A⇓ ; and see Fig. 1 in the Data Supplement that accompanies the online version of this Letter at http://www.clinchem.org/content/vol53/issue1). Figure 1. Principle ( A ) and standard curves ( B ) of the upconversion FRET-based competitive homogeneous E2 immunoassay. ( A ), the anti-E2-Fab-coated FCD-546–1 UCP (donor) can bind E2 (analyte) and E2-AF680 conjugate (acceptor). Phosphor emission at … |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|