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Screening Scheme Based on Measurement of Fluorescence Lifetime in the Nanosecond Domain
1 University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, Regensburg, Germany.
* To whom correspondence should be addressed. E-mail: otto.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
First published on August 29, 2005, doi:10.1177/1087057105277493 This article has been cited by other articles:
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