WO2004034054A3 - Method for enhancing and prolonging the bioluminescence resonance energy transfer (bret) signal in a bret assay and a substrate solution for use in a bret assay - Google Patents

Method for enhancing and prolonging the bioluminescence resonance energy transfer (bret) signal in a bret assay and a substrate solution for use in a bret assay Download PDF

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Publication number
WO2004034054A3
WO2004034054A3 PCT/DK2003/000687 DK0300687W WO2004034054A3 WO 2004034054 A3 WO2004034054 A3 WO 2004034054A3 DK 0300687 W DK0300687 W DK 0300687W WO 2004034054 A3 WO2004034054 A3 WO 2004034054A3
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WO
WIPO (PCT)
Prior art keywords
bret
arrestin
assay
fusion protein
signal
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/DK2003/000687
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French (fr)
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WO2004034054A2 (en
Inventor
Anders Heding
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
7TM Pharma AS
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7TM Pharma AS
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Filing date
Publication date
Application filed by 7TM Pharma AS filed Critical 7TM Pharma AS
Priority to US10/531,072 priority Critical patent/US20060099646A1/en
Priority to AU2003300424A priority patent/AU2003300424A1/en
Priority to EP03756450A priority patent/EP1554581A2/en
Publication of WO2004034054A2 publication Critical patent/WO2004034054A2/en
Publication of WO2004034054A3 publication Critical patent/WO2004034054A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
    • G01N2333/726—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Abstract

An improved BRET assay, wherein the BRET signal is enhanced and/or prolonged. The improved BRET assay comprises the steps of i) adding a substrate to a cell comprising GPCR-Rluc fusion protein and a β -arrestin-GFP fusion protein, wherein the (β-arrestin is mutated, ii) adding a ligand to obtain, if possible, a GPCR-Rluc/(β-arrestin-GFP complex, and iii) measuring a BRET signal to obtain a BRET ratio, wherein the improvement leads to an increased BRET ratio compared with the ratios obtained by use of the same process employing a β-arrestin-GFP fusion protein wherein the β-arrestin is the wild type β-arrestin, or employing a 13-arrestin-GFP fusion protein, wherein the (β-arrestin is a β-arrestin specifically mutated so that it acts on the receptor independent of the receptors phosphorylation state. The invention further relates to a stable substrate solution for use in an improved BRET assay.
PCT/DK2003/000687 2002-10-11 2003-10-10 Method for enhancing and prolonging the bioluminescence resonance energy transfer (bret) signal in a bret assay and a substrate solution for use in a bret assay Ceased WO2004034054A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US10/531,072 US20060099646A1 (en) 2002-10-11 2003-10-10 Bret assay
AU2003300424A AU2003300424A1 (en) 2002-10-11 2003-10-10 Method for enhancing and prolonging the bioluminescence resonance energy transfer (bret) signal in a bret assay and a substrate solution for use in a bret assay
EP03756450A EP1554581A2 (en) 2002-10-11 2003-10-10 Method for enhancing and prolonging the bioluminescence resonance energy transfer (bret) signal in a bret assay and a substrate solution for use in a bret assay

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DKPA200201568 2002-10-11
DKPA200201568 2002-10-11
DKPA200300076 2003-01-22
DKPA200300076 2003-01-22

Publications (2)

Publication Number Publication Date
WO2004034054A2 WO2004034054A2 (en) 2004-04-22
WO2004034054A3 true WO2004034054A3 (en) 2004-07-29

Family

ID=32094937

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK2003/000687 Ceased WO2004034054A2 (en) 2002-10-11 2003-10-10 Method for enhancing and prolonging the bioluminescence resonance energy transfer (bret) signal in a bret assay and a substrate solution for use in a bret assay

Country Status (4)

Country Link
US (1) US20060099646A1 (en)
EP (1) EP1554581A2 (en)
AU (1) AU2003300424A1 (en)
WO (1) WO2004034054A2 (en)

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK1888788T3 (en) * 2005-06-03 2011-11-28 Beacon Biotechnology Llc Chemiluminescence nucleic acid assay
US9091659B2 (en) 2006-10-17 2015-07-28 Zymera, Inc. Hydrolase detection system with sterically caged substrates
WO2008049036A2 (en) * 2006-10-17 2008-04-24 Zymera, Inc. Enzyme detection system with caged substrates
WO2008109459A1 (en) 2007-03-02 2008-09-12 University Of Massachusetts Luciferins
CA2748500C (en) 2009-01-29 2017-10-17 Commonwealth Scientific And Industrial Research Organisation Measuring g protein coupled receptor activation
US8524457B2 (en) 2009-09-22 2013-09-03 William Patterson Method for the selection of specific affinity binders by homogeneous noncompetitive assay
US8795977B2 (en) 2009-12-02 2014-08-05 Conopco, Inc. Method for screening a potential modulator compound of a taste receptor
US10024862B2 (en) * 2012-12-12 2018-07-17 Promega Corporation Recognition of cellular target binding by a bioactive agent using intracellular bioluminescence resonance energy transfer
US10067149B2 (en) 2012-12-12 2018-09-04 Promega Corporation Recognition of cellular target binding by a bioactive agent using intracellular bioluminescence resonance energy transfer
EP2999700A4 (en) * 2013-05-20 2017-02-22 The Regents of The University of California Genetically encoded sensors for imaging proteins and their complexes
AU2015233370A1 (en) * 2014-03-20 2016-09-22 Centre National De La Recherche Scientifique (Cnrs) Use of compounds inhibiting Apelin / APJ / GP130 signaling for treating cancer

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5891646A (en) * 1997-06-05 1999-04-06 Duke University Methods of assaying receptor activity and constructs useful in such methods
WO2001067106A2 (en) * 2000-03-03 2001-09-13 Millennium Pharmaceuticals, Inc. Identification of g protein-coupled receptor ligands and modulators
WO2002046763A1 (en) * 2000-12-04 2002-06-13 Novo Nordisk A/S Method of identifying compounds capable of acting as agonists or antagonists of g-protein coupled receptors

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030203404A1 (en) * 1999-12-22 2003-10-30 Erik Joly Bioluminescence resonance energy transfer( bret) system with broad spectral resolution between donor and acceptor emission wavelengths and its use
US6893827B1 (en) * 2000-02-07 2005-05-17 Applera Corporation Receptor function assay for G-protein coupled receptors and orphan receptors by reporter enzyme mutant complementation
US20030157553A1 (en) * 2000-03-03 2003-08-21 Gabriel Berstein Methods of assaying for G protein-coupled receptor ligands and modulators
WO2003097795A2 (en) * 2002-05-13 2003-11-27 Norak Biosciences, Inc. Constitutively translocating cell line

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5891646A (en) * 1997-06-05 1999-04-06 Duke University Methods of assaying receptor activity and constructs useful in such methods
WO2001067106A2 (en) * 2000-03-03 2001-09-13 Millennium Pharmaceuticals, Inc. Identification of g protein-coupled receptor ligands and modulators
WO2002046763A1 (en) * 2000-12-04 2002-06-13 Novo Nordisk A/S Method of identifying compounds capable of acting as agonists or antagonists of g-protein coupled receptors

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ABRAHAM KOVOOR ET AL: "Targeted Construction of Phosphorylation-independent Beta-Arrestin Mutants with Constitutive Activity in Cells", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 274, no. 11, 12 March 1999 (1999-03-12), pages 6831 - 6834, XP002267973 *
ERIK JOLY ET AL: "Bioluminescence Resonance Energy Transfer (BRET 2 TM). Principle, Applications and Products", PACKARD BIOSCIENCE, BIOSIGNAL PACKARD INC., 2001, pages 1 - 6, XP002267972, Retrieved from the Internet <URL:http://www2.cbm.uam.es/confocal/Manuales/bret.pdf> [retrieved on 20040126] *

Also Published As

Publication number Publication date
EP1554581A2 (en) 2005-07-20
AU2003300424A1 (en) 2004-05-04
US20060099646A1 (en) 2006-05-11
WO2004034054A2 (en) 2004-04-22
AU2003300424A8 (en) 2004-05-04

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