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Screening Scheme Based on Measurement of Fluorescence Lifetime in the Nanosecond DomainUniversity of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, Regensburg, Germany
Innovative Optische Messtechnik GmbH, Berlin, Germany
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
This version was published on October
1, 2005 Journal of Biomolecular Screening, Vol. 10, No. 7,
687-694 (2005) This article has been cited by other articles:
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