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This version was published on October 1, 2005
Journal of Biomolecular Screening, Vol. 10, No. 7, 687-694 (2005)
DOI: 10.1177/1087057105277493

Screening Scheme Based on Measurement of Fluorescence Lifetime in the Nanosecond Domain

Bianca K. Hoefelschweiger

University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, Regensburg, Germany

Lutz Pfeifer

Innovative Optische Messtechnik GmbH, Berlin, Germany

Otto S. Wolfbeis

University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, Regensburg, Germany; University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germanyotto.wolfbeis{at}chemie.uni-regensburg.de

The authors demonstrate that the fluorescence lifetime of certain fluorescent labels is a useful parameter to detect affinity binding between biotin and streptavidin, as well as between biotinylated bovine serum albumin and streptavidin. The assay is performed in a microplate format, and lifetimes are determined using dye laser-induced fluorescence. Four fluorescent labels are presented that undergo a significant change in their lifetime upon affinity binding. The scheme, referred to as the fluorescence lifetime affinity assay, has several attractive features in that it requires single labeling only, represents a homogeneous assay, allows each of the 2 binding partners to be labeled, and is compatible with the standard microwell formats used in high-throughput screening.

Key Words: Laser-induced fluorescence • lifetime • fluorescent label • affinity binding • affinity assay • decay time


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