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Terbium to quantum dot FRET Bioconjugates for clinical diagnostics influence of human plasma on optical and assembly properties

  • Forster resonance energy transfer (FRET) from luminescent terbium complexes (LTC) as donors to semiconductor quantum dots (QDs) as acceptors allows extraordinary large FRET efficiencies due to the long Forster distances afforded. Moreover, time-gated detection permits an efficient suppression of autofluorescent background leading to sub-picomolar detection limits even within multiplexed detection formats. These characteristics make FRET-systems with LTC and QDs excellent candidates for clinical diagnostics. So far, such proofs of principle for highly sensitive multiplexed biosensing have only been performed under optimized buffer conditions and interactions between real-life clinical media such as human serum or plasma and LTC-QD-FRET-systems have not yet been taken into account. Here we present an extensive spectroscopic analysis of absorption, excitation and emission spectra along with the luminescence decay times of both the single components as well as the assembled FRET-systems in TRIS-buffer, TRIS-buffer with 2% bovine serumForster resonance energy transfer (FRET) from luminescent terbium complexes (LTC) as donors to semiconductor quantum dots (QDs) as acceptors allows extraordinary large FRET efficiencies due to the long Forster distances afforded. Moreover, time-gated detection permits an efficient suppression of autofluorescent background leading to sub-picomolar detection limits even within multiplexed detection formats. These characteristics make FRET-systems with LTC and QDs excellent candidates for clinical diagnostics. So far, such proofs of principle for highly sensitive multiplexed biosensing have only been performed under optimized buffer conditions and interactions between real-life clinical media such as human serum or plasma and LTC-QD-FRET-systems have not yet been taken into account. Here we present an extensive spectroscopic analysis of absorption, excitation and emission spectra along with the luminescence decay times of both the single components as well as the assembled FRET-systems in TRIS-buffer, TRIS-buffer with 2% bovine serum albumin, and fresh human plasma. Moreover, we evaluated homogeneous LTC-QD FRET assays in QD conjugates assembled with either the well-known, specific biotin-streptavidin biological interaction or, alternatively, the metal-affinity coordination of histidine to zinc. In the case of conjugates assembled with biotin-streptavidin no significant interference with the optical and binding properties occurs whereas the histidine-zinc system appears to be affected by human plasma.show moreshow less

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Author details:Frank Morgner, Stefan Stufler, Daniel Geissler, Igor L. Medintz, W. Russ Algar, Kimihiro Susumu, Michael H. Stewart, Juan B. Blanco-Canosa, Philip E. Dawson, Niko HildebrandtGND
DOI:https://doi.org/10.3390/s111009667
ISSN:1424-8220
Title of parent work (English):Sensors
Publisher:MDPI
Place of publishing:Basel
Publication type:Article
Language:English
Year of first publication:2011
Publication year:2011
Release date:2017/03/26
Tag:FRET; biotin; blood; clinical diagnostics; histidin; immunoassay; luminescence lifetime; plasma; quantum dots; streptavidin; terbium
Volume:11
Issue:10
Number of pages:18
First page:9667
Last Page:9684
Funding institution:DTRA; DARPA; ONR; NRL; German Bundesministerium fur Bildung und Forschung; European Commission; Innovative Medicines Initiative
Organizational units:Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie
Peer review:Referiert
Publishing method:Open Access
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