WO2002085667A3 - Three cube fret method (33-fret) for detecting fluorescence energy transfer - Google Patents

Three cube fret method (33-fret) for detecting fluorescence energy transfer Download PDF

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Publication number
WO2002085667A3
WO2002085667A3 PCT/US2002/004563 US0204563W WO02085667A3 WO 2002085667 A3 WO2002085667 A3 WO 2002085667A3 US 0204563 W US0204563 W US 0204563W WO 02085667 A3 WO02085667 A3 WO 02085667A3
Authority
WO
WIPO (PCT)
Prior art keywords
fret
interactions
tagged molecules
acceptor
donor
Prior art date
Application number
PCT/US2002/004563
Other languages
French (fr)
Other versions
WO2002085667A2 (en
Inventor
David T Yue
Michael G Erickson
Original Assignee
Univ Johns Hopkins Med
David T Yue
Michael G Erickson
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Univ Johns Hopkins Med, David T Yue, Michael G Erickson filed Critical Univ Johns Hopkins Med
Priority to AU2002338407A priority Critical patent/AU2002338407A1/en
Priority to US10/467,964 priority patent/US20040191786A1/en
Publication of WO2002085667A2 publication Critical patent/WO2002085667A2/en
Publication of WO2002085667A3 publication Critical patent/WO2002085667A3/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)

Abstract

The invention provides a method for determining a measure of FRET comprising obtaining sequential fluorescent intensity readings from a specimen, such as a cell using three filter cubes. Simple equations manipulate readings from each of the filter sets or cubes to specify a unitless index of FRET called the FRET ratio (FR). FR bears a linear relation to FRET efficiency E. The method also provides for determining the fraction of acceptor-tagged molecules bound by donor-tagged molecules; the relative affinity of binding; and the strength of FRET interactions when all acceptor-tagged molecules are bound by donor. The latter determination enables estimates of the physical distance and/or orientation between interacting fluorophore molecules. The method can be used to detect analytes or inter- or intra-molecular interactions. In a preferred aspect, the method is used in an HTS assay to identify modulators of such interactions.
PCT/US2002/004563 2001-02-16 2002-02-15 Three cube fret method (33-fret) for detecting fluorescence energy transfer WO2002085667A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2002338407A AU2002338407A1 (en) 2001-02-16 2002-02-15 Three cube fret method (33-fret) for detecting fluorescence energy transfer
US10/467,964 US20040191786A1 (en) 2001-02-16 2002-02-15 Three cube fret method (3-fret) for detecting fluorescence energy transfer

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US26966901P 2001-02-16 2001-02-16
US60/269,669 2001-02-16
US27591101P 2001-03-15 2001-03-15
US60/275,911 2001-03-15

Publications (2)

Publication Number Publication Date
WO2002085667A2 WO2002085667A2 (en) 2002-10-31
WO2002085667A3 true WO2002085667A3 (en) 2003-02-06

Family

ID=26953827

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/004563 WO2002085667A2 (en) 2001-02-16 2002-02-15 Three cube fret method (33-fret) for detecting fluorescence energy transfer

Country Status (2)

Country Link
AU (1) AU2002338407A1 (en)
WO (1) WO2002085667A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FI20070163A0 (en) 2007-02-27 2007-02-27 Hidex Oy Improved homogeneous luminescence bioassay
CN113899722B (en) * 2021-09-14 2022-11-01 华南师范大学 Method for measuring FRET system correction parameter based on single standard FRET plasmid and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6171780B1 (en) * 1997-06-02 2001-01-09 Aurora Biosciences Corporation Low fluorescence assay platforms and related methods for drug discovery
US6200762B1 (en) * 1997-08-01 2001-03-13 Aurora Biosciences Corporation Photon reducing agents and compositions for fluorescence assays
US6221612B1 (en) * 1997-08-01 2001-04-24 Aurora Biosciences Corporation Photon reducing agents for use in fluorescence assays
US6323331B1 (en) * 1996-09-13 2001-11-27 The Trustees Of Columbia University In The City Of New York Beclin-related nucleic acid molecules, and uses thereof
US6329139B1 (en) * 1995-04-25 2001-12-11 Discovery Partners International Automated sorting system for matrices with memory

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6329139B1 (en) * 1995-04-25 2001-12-11 Discovery Partners International Automated sorting system for matrices with memory
US6323331B1 (en) * 1996-09-13 2001-11-27 The Trustees Of Columbia University In The City Of New York Beclin-related nucleic acid molecules, and uses thereof
US6171780B1 (en) * 1997-06-02 2001-01-09 Aurora Biosciences Corporation Low fluorescence assay platforms and related methods for drug discovery
US6200762B1 (en) * 1997-08-01 2001-03-13 Aurora Biosciences Corporation Photon reducing agents and compositions for fluorescence assays
US6221612B1 (en) * 1997-08-01 2001-04-24 Aurora Biosciences Corporation Photon reducing agents for use in fluorescence assays

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SONG ET AL.: "Flow cytometry-based biosensor for detection of multivalent proteins", ANALYTICAL BIOCHEMISTRY, vol. 284, no. 1, 15 August 2000 (2000-08-15), pages 35 - 41, XP002955981 *

Also Published As

Publication number Publication date
AU2002338407A1 (en) 2002-11-05
WO2002085667A2 (en) 2002-10-31

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