Screening scheme based on measurement of fluorescence lifetime in the nanosecond domain.
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.
Hoefelschweiger
Bianca K
BK
University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, Regensburg, Germany.
Pfeifer
Lutz
L
Wolfbeis
Otto S
OS
D000345:Affinity Labels; D001710:Biotin / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D019809:Streptavidin / Q000378:metabolism*; D013997:Time Factors
D000345:Affinity Labels; D005456:Fluorescent Dyes; D001710:Biotin; D019809:Streptavidin