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Augustin, Christine M. ; Oswald, Bernhard ; Wolfbeis, Otto S.

Time-Resolved Luminescence Energy Transfer Immunobinding Study Using a Ruthenium–Ligand Complex as a Donor Label

Augustin, Christine M., Oswald, Bernhard and Wolfbeis, Otto S. (2002) Time-Resolved Luminescence Energy Transfer Immunobinding Study Using a Ruthenium–Ligand Complex as a Donor Label. Analytical Biochemistry 305 (2), pp. 166-172.

Date of publication of this fulltext: 13 May 2011 08:27
Article
DOI to cite this document: 10.5283/epub.20748


Abstract

A novel immunosystem is described that exploits the effect of luminescence energy transfer from a luminescently labeled antigen to a fluorescent antibody. A luminescent ruthenium-ligand complex (D-455) with absorption/emission maxima at 456/639 nm, respectively, was employed as the donor label, and a squaraine-type cyanine label (636/655 nm), as the fluorescent acceptor label. Specifically, the ...

A novel immunosystem is described that exploits the effect of luminescence energy transfer from a luminescently labeled antigen to a fluorescent antibody. A luminescent ruthenium-ligand complex (D-455) with absorption/emission maxima at 456/639 nm, respectively, was employed as the donor label, and a squaraine-type cyanine label (636/655 nm), as the fluorescent acceptor label. Specifically, the system human serum albumin (HSA)/anti-HSA was studied. HSA was labeled with the donor dye D-455, and anti-HSA was labeled with the acceptor dye A-631. On formation of the antigen-antibody complex, energy transfer occurs. The radiationless energy transfer affects both the decay time of D-455 and the intensities of the emissions of both D-455 and A-631. The decay time of around 500 ns of D-455 allows frequency-domain measurements in the low kilohertz range and therefore can be based on the use of conventional optoelectronics. This also suggests gated measurements to be performed. The major difference from existing HSA immunosystems is the use of a slow decaying ruthenium-ligand complex as the donor and of a long-wave emitting cyanine acceptor dye having a high quantum yield and a decay kinetics that is governed by the rate of energy transfer from the slow decaying donor. (C) 2002 Elsevier Science (USA).



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Details

Item typeArticle
Journal or Publication TitleAnalytical Biochemistry
Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE
Place of Publication:SAN DIEGO
Volume:305
Number of Issue or Book Chapter:2
Page Range:pp. 166-172
Date2002
InstitutionsChemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Antje J. Bäumner, formerly Prof. Wolfbeis)
Identification Number
ValueType
10.1006/abio.2002.5633DOI
KeywordsTRANSFER IMMUNOASSAY; FLUORESCENCE; luminescence immunoassay; time resolution; luminescence decay time; resonance energy transfer (RET); fluorescent labels; ruthenium labels
Dewey Decimal Classification500 Science > 540 Chemistry & allied sciences
StatusPublished
RefereedUnknown
Created at the University of RegensburgUnknown
Item ID20748

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