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.
Liu B, Li S, Hu J: Technological advances in high-throughput screening. Am J PharmacoGenomics2004;4:263-276.
2.
Wilson WD: Analyzing biomolecular interactions. Science2002;295:2103-2105.
3.
Doman TN, McGovern SL, Witherbee BJ, Kasten TP, Stallings WC, Connolly DT, et al: Molecular docking and high-throughput screening for novel inhibitors of protein tyrosine phosphatase-1B. J Med Chem2002;45:2213-2221.
4.
Sills MA, Weiss D, Pham Q, Schweitzer R, Wu X, Wu JJ: Comparison of assay technologies for a tyrosine kinase assay generates different results in high throughput screening. J Biomol Screen2002;7:191-214.
5.
Steff AM, Fortin M, Arguin C, Hugo P: Detection of a decrease in green fluorescent protein fluorescence for the monitoring of cell death: an assay amenable to high-throughput screening technologies. Cytometry2001;45:237-243.
6.
Owicki JC: Fluorescence polarization and anisotropy in high throughput screening: perspectives and primer. J Biomol Screen2000;5:297-306.
7.
Medina MA, Schwille P: Fluorescence correlation spectroscopy for the detection and study of single molecules in biology. BioEssays2002;24:758-764.
8.
Roda A, Guardigli M, Pasini P, Mirasoli M: Bioluminescence and chemiluminescence in drug screening. Anal Bioanal Chem2003;377:826-833.
9.
Haupt K: Imprinted polymers: tailor-made mimics of antibodies and receptors. Chem Comm2003;Jan 21(2):171-178.
10.
Senholdt M, Siemann M, Mosbach K, Andersson LI: Determination of cyclosporin A and metabolites total concentration using a molecularly imprinted polymer based radioligand binding assay. Anal Lett1997;30:1809-1821.
11.
Wijayawardhana CA, Halsall HB, Heineman WR: Electrochemical immunoassay. In Bard AJ, Stratmann M (eds): Encyclopedia of Electrochemistry: Vol. 9. Bioelectrochemistry. Weinheim: Wiley-VCH, 2002: 145, 147-174.
12.
Stefan RI, van Staden JF, Aboul Enein HY: Design and use of electrochemical sensors in enantioselective high throughput screening of drugs: a minireview. Combinat Chem High Throughput Screen2000;3:445-454.
13.
Kerns EH: High throughput physicochemical profiling for drug discovery. J Pharm Sci2001;90:1838-1858.
14.
Huber C, Klimant I, Krause Ch, Wolfbeis OS: Dual lifetime referencing as applied to a chloride optical sensor. Anal Chem2001;73:2097-2103.
15.
Valeur B: Molecular Fluorescence, Principles and Applications. Weinheim: Wiley-VCH, 2002.
16.
Liebsch G, Klimant I, Frank B, Holst G, Wolfbeis OS: Luminescence lifetime imaging of oxygen, pH, and carbon dioxide distribution using optical sensors. Appl Spectrosc2000;54:548-559.
17.
Hemmilä I, Webb S: Time-resolved fluorometry: an overview of the labels and core technologies for drug screening applications. Drug Discovery Today1997;2:373-381.
18.
Mathis G: Rare earth cryptates and homogeneous fluoroimmunoassays with human sera. Clin Chem1993;39:1953-1959.
Duerkop A, Lehmann F, Wolfbeis OS: Polarization immunoassays using reactive ruthenium metal-ligand complexes as luminescent labels. Anal Bioanal Chem2002;372:688-694.
21.
Augustin CM, Oswald B, Wolfbeis OS: Time-resolved luminescence energy transfer immunobinding study using a ruthenium-ligand complex as a donor label. Anal Biochem2002;305:166-172.
22.
Jäger S, Brand L, Eggeling C: New fluorescence techniques for high-throughput drug discovery. Curr Pharm Biotechnol2003;4:463-476.
23.
Eggeling Ch, Brand L, Ullmann D, Jäger S: Highly sensitive fluorescence detection technology currently available for HTS. Drug Discovery Today2003;8:632-641.
24.
French TE, Bailey B, Stumbo DP, Modlin DN: Time-resolved fluorometer for high-throughput screening. Proc Soc Photoinstrum Eng (SPIE)1999;3603:272-280.
25.
Turconi S, Bingham RP, Haupts U, Pope AJ: Developments in fluorescence lifetime-based analysis for ultra-HTS. Drug Discovery Today2001;6:S27-S39.
26.
Gruber M, Oswald B, Wetzl B, Enderlein J, Wolfbeis OS: Long wavelength absorbing fluorophores and their application to oligonucleotide labeling and fluorescence resonance energy transfer (FRET) hybridization studies. JFluorescence2005;15:2007-2014.
27.
Wetzl BK, Yarmoluk SM, Craig DB, Wolfbeis OS: Chameleon labels for staining and quantifying of proteins. Angew Chem Int Ed2004;43:5400-5402.
28.
Kostenko OM, Dmitrieva SY, Tolmachev OI, Yarmoluk SM: New approach for fluorescent peptide labeling with pyrylium cyanine dyes. J Fluoresc2002;12:173-175.
29.
Huang S, Stump MD, Weiss R, Caldwell KD: Binding of biotinylated DNA to streptavidin-coated polystyrene latex: effects of chain length and particle size. Anal Biochem1996;237:115-122.
30.
Diamandis EP, Christopoulos TK: Europium chelate labels in time-resolved immunoassays and DNA hybridization assays. Anal Chem1990;62:1149A-1157A.
31.
Zhang JH, Chung TD, Oldenburg KR: A simple statistical parameter for use in evaluation and validation of high-throughput screening assays. J Biomol Screen1999;4:67-73.
