Title:
METHODS OF SCREENING FOR LIGANDS OF TARGET MOLECULES
Document Type and Number:
WIPO Patent Application WO2002103321
Kind Code:
A3
Abstract:
The present invention provides methods of screening for ligands of target molecules. The methods of the present invention include assays in which a target molecule is subjected to denaturing conditions, and compounds are screened for the ability to alter the susceptibility of the target to unfolding. The methods of the present invention use fluorescence detection to determine that degree of unfolding of a target molecule. In some aspects of the present invention, fluorescence resonance energy transfer (FRET) is detected. In other aspects of the invention, fluorescence polarization (FP) is detected. In preferred embodiments, a target molecule such as a target protein is heated to a temperature, called TATLAS, at which at least a portion of the target molecule unfolds, in the presence of a test compound. In some embodiments of the present invention, the degree of unfolding of the target molecule is determined by binding of a specific binding member specific for the unfolded form of a target molecule that is coupled to a fluorophore that can participate in FRET. In some other embodiments of the present invention, the degree of unfolding of a target molecule is determined by FRET detection of aggregates of the target molecule. In yet other embodiments of the present invention, the degree of unfolding of a target molecule is determined by detection of fluorescence polarization of aggregates of the target molecule. The present invention provides sensitive, high throughput screens for identifying ligands of target molecules that are not dependent on the identity or function of the target.
Inventors:
DJABALLAH HAKIM
RONGEY SCOTT
PATEL RUPAL
WANG MEI MEI
COYLE JOSEPH
LI BIN
WORLAND STEPHEN
RONGEY SCOTT
PATEL RUPAL
WANG MEI MEI
COYLE JOSEPH
LI BIN
WORLAND STEPHEN
Application Number:
PCT/US2002/018952
Publication Date:
March 20, 2003
Filing Date:
June 13, 2002
Export Citation:
Assignee:
ANADYS PHARMACEUTICALS INC (US)
International Classes:
G01N21/78; G01N33/15; G01N33/50; G01N33/53; G01N33/536; G01N33/542; G01N33/566; (IPC1-7): G01N33/573; C12N15/62
Domestic Patent References:
| WO2002033102A1 | 2002-04-25 |
Foreign References:
| US6399392B1 | 2002-06-04 | |||
| US6342379B1 | 2002-01-29 | |||
| US6465199B1 | 2002-10-15 | |||
| US6451543B1 | 2002-09-17 |
Other References:
See also references of EP 1409982A4
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