DE865502T1 - FLUORESCENT-BASED DETECTION METHOD FOR LIGANDS - Google Patents

FLUORESCENT-BASED DETECTION METHOD FOR LIGANDS

Info

Publication number
DE865502T1
DE865502T1 DE0865502T DE96944304T DE865502T1 DE 865502 T1 DE865502 T1 DE 865502T1 DE 0865502 T DE0865502 T DE 0865502T DE 96944304 T DE96944304 T DE 96944304T DE 865502 T1 DE865502 T1 DE 865502T1
Authority
DE
Germany
Prior art keywords
test
target protein
ligand
control compositions
protein
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.)
Pending
Application number
DE0865502T
Other languages
German (de)
Inventor
Andrew Pakula
Robert Sauer
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.)
Scriptgen Pharmaceuticals Inc
Original Assignee
Scriptgen Pharmaceuticals Inc
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 Scriptgen Pharmaceuticals Inc filed Critical Scriptgen Pharmaceuticals Inc
Publication of DE865502T1 publication Critical patent/DE865502T1/en
Pending legal-status Critical Current

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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6811—Selection methods for production or design of target specific oligonucleotides or binding molecules
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • 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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pathology (AREA)
  • Biophysics (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Peptides Or Proteins (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Claims (8)

DE/EP O 885 502 T1 Europäische Patentanmeldung Nr.: 96 944 304.3 Anmelder SCRIPTGEN PHARMACEUTICALS, INC. PatentansprücheDE/EP O 885 502 T1 European patent application No: 96 944 304.3 Applicant SCRIPTGEN PHARMACEUTICALS, INC. Patent claims 1. Auf Fluoreszenz basierendes Absuchverfahren zur Identifizierung eines Liganden, der an ein vorher bestimmtes Zielprotein bindet, wobei das Verfahren die folgenden Schritte umfaßt:1. A fluorescence-based screening method for identifying a ligand that binds to a predetermined target protein, the method comprising the following steps: (a) Auswahl einer großen Anzahl von Verbindungen als Testliganden, von denen nicht bekannt ist, ob sie an das Zielprotein binden;(a) selection of a large number of compounds as test ligands, for which it is not known whether they bind to the target protein; (b) Inkubation des Zielproteins zusammen mit jedem der Testliganden, wodurch eine Testzusammensetzung hergestellt wird, und in Abwesenheit eines Testliganden, wodurch eine Kontrollzusammensetzung hergestellt wird;(b) incubating the target protein together with each of the test ligands, thereby producing a test composition, and in the absence of a test ligand, thereby producing a control composition; (c) Inkontaktbringen der Test- und Kontrollzusammensetzungen mit einer Konformation-sensitiven Fluoreszenzsonde;(c) contacting the test and control compositions with a conformation-sensitive fluorescent probe; (d) Behandlung der Test- und Kontrollzusammensetzungen unter Bedingungen, die bewirken, daß das Zielprotein bis zu einem geeigneten Ausmaß entfaltet wird;(d) treating the test and control compositions under conditions effective to unfold the target protein to an appropriate extent; (e) Messung der Fluoreszenzemission der Konformation-sensitiven Fluoreszenzsonde in den Test- und Kontrollzusammensetzungen; und(e) measuring the fluorescence emission of the conformation-sensitive fluorescent probe in the test and control compositions; and (f) Vergleich der in Schritt (d) durchgeführten Messung der Test- und Kontrollzusammensetzungen, wobei der Testligand ein Ligand ist, der an das Zielprotein bindet, wenn die Intensität der Fluoreszenzemission der Sonde in der Testzusammensetzung stärker oder schwächer ist als in der Kontrollzusammensetzung. '(f) comparing the measurement of the test and control compositions made in step (d), wherein the test ligand is a ligand that binds to the target protein when the intensity of the fluorescence emission of the probe is stronger or weaker in the test composition than in the control composition. ' 2. Verfahren nach Anspruch 1, das außerdem die Wiederholung der Schritte (b) bis (f) mit einer großen Anzahl von Testliganden umfaßt, bis ein Ligand identifiziert ist, der an das Zielprotein bindet.2. The method of claim 1, further comprising repeating steps (b) to (f) with a large number of test ligands until a ligand is identified that binds to the target protein. 3. Verfahren nach Anspruch 1, wobei die Fluoreszenzsonde vorzugsweise an die gefaltete, ungefaltete oder geschmolzene globuläre Form des Proteins bindet.3. The method of claim 1, wherein the fluorescent probe preferentially binds to the folded, unfolded or molten globular form of the protein. ..... .p.E/EP O 365502 Tl..... .p.E/EP O 365502 Tl 4. Verfahren nach Anspruch 3, wobei die Sonde aus der Gruppe ausgewählt ist, die aus 1-Anilino-8-naphthalensulfonat (ANS), Bis-1-anilino-8-naphthalensulfonat (Bis-ANS) und 6-Propionyl-2-(N,N-dimethyl)aminonaphthalen (Prodan) besteht. 4. The method of claim 3, wherein the probe is selected from the group consisting of 1-anilino-8-naphthalenesulfonate (ANS), bis-1-anilino-8-naphthalenesulfonate (bis-ANS) and 6-propionyl-2-(N,N-dimethyl)aminonaphthalene (prodan). 5. Verfahren nach Anspruch 4, wobei die Fluoreszenzsonde Bis-ANS ist.5. The method of claim 4, wherein the fluorescent probe is Bis-ANS. 6. Verfahren nach Anspruch 1, wobei die Behandlung umfaßt, daß die Temperatur, der die Test- und Kontrollzusammensetzungen ausgesetzt werden, erhöht wird, daß die Test- und Kontrollzusammensetzungen mit einem Protein-Denaturierungsmittel in Kontakt gebracht werden oder daß Kombinationen davon angewendet werden.6. The method of claim 1, wherein the treatment comprises increasing the temperature to which the test and control compositions are exposed, contacting the test and control compositions with a protein denaturant, or combinations thereof. 7. Verfahren nach Anspruch 1, wobei das Zielprotein im Vergleich zur Wildtyp-Version des Proteins stabilisierende oder destabilisierende Mutationen enthält.7. The method of claim 1, wherein the target protein contains stabilizing or destabilizing mutations compared to the wild-type version of the protein. 8. Verfahren nach Anspruch 1, wobei der Testligand aus der Gruppe ausgewählt ist, die aus Metallen, Peptiden, Proteinen, Lipiden, Polysacchariden, Nucleinsäuren, kleinen organischen Molekülen und Kombinationen davon besteht.8. The method of claim 1, wherein the test ligand is selected from the group consisting of metals, peptides, proteins, lipids, polysaccharides, nucleic acids, small organic molecules, and combinations thereof.
DE0865502T 1995-12-07 1996-12-06 FLUORESCENT-BASED DETECTION METHOD FOR LIGANDS Pending DE865502T1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US834695P 1995-12-07 1995-12-07
PCT/US1996/019698 WO1997020952A1 (en) 1995-12-07 1996-12-06 A fluorescence-based screening method for identifying ligands

Publications (1)

Publication Number Publication Date
DE865502T1 true DE865502T1 (en) 1999-08-19

Family

ID=21731102

Family Applications (1)

Application Number Title Priority Date Filing Date
DE0865502T Pending DE865502T1 (en) 1995-12-07 1996-12-06 FLUORESCENT-BASED DETECTION METHOD FOR LIGANDS

Country Status (5)

Country Link
EP (1) EP0865502A4 (en)
JP (1) JP2000502440A (en)
CA (1) CA2236339A1 (en)
DE (1) DE865502T1 (en)
WO (1) WO1997020952A1 (en)

Families Citing this family (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2184195C (en) 1995-10-25 2002-04-16 Andrew Pakula Screening method for identifying ligands for target proteins
EP0914608A1 (en) 1996-05-09 1999-05-12 3-Dimensional Pharmaceuticals, Inc. Microplate thermal shift assay and apparatus for ligand development and multi-variable protein chemistry optimization
JP2001503546A (en) 1996-11-04 2001-03-13 ３―ディメンショナル ファーマシューティカルズ インコーポレイテッド Systems, methods, and computer program products for visualizing and interactively processing and analyzing chemical data
US6453246B1 (en) 1996-11-04 2002-09-17 3-Dimensional Pharmaceuticals, Inc. System, method, and computer program product for representing proximity data in a multi-dimensional space
US6571227B1 (en) 1996-11-04 2003-05-27 3-Dimensional Pharmaceuticals, Inc. Method, system and computer program product for non-linear mapping of multi-dimensional data
US6261842B1 (en) 1997-10-23 2001-07-17 Wisconsin Alumni Research Foundation Microorganism genomics, compositions and methods related thereto
IL136046A0 (en) * 1997-11-12 2001-05-20 Dimensional Pharm Inc Methods for classifying proteins
US20020010550A1 (en) 1998-09-14 2002-01-24 George M. Grass Pharmacokinetic-based drug design tool and method
US6569631B1 (en) 1998-11-12 2003-05-27 3-Dimensional Pharmaceuticals, Inc. Microplate thermal shift assay for ligand development using 5-(4″dimethylaminophenyl)-2-(4′-phenyl)oxazole derivative fluorescent dyes
US6358708B1 (en) 1999-02-04 2002-03-19 Anadys Pharmaceuticals, Inc. Candida albicans SRB-7
US6972198B2 (en) 1999-02-26 2005-12-06 Cyclacel, Ltd. Methods and compositions using protein binding partners
US6376180B1 (en) * 1999-12-09 2002-04-23 Pharmacia & Upjohn Company Methods of identifying compounds that bind to target species under isothermal denaturing conditions
US6582907B1 (en) 1999-12-09 2003-06-24 Pharmacia & Upjohn Company Use of fluorescence correlation spectroscopy to identify compounds that bind to target species under isothermal denaturing conditions
US7416524B1 (en) 2000-02-18 2008-08-26 Johnson & Johnson Pharmaceutical Research & Development, L.L.C. System, method and computer program product for fast and efficient searching of large chemical libraries
US6671627B2 (en) 2000-02-29 2003-12-30 3-D Pharmaceuticals, Inc. Method and computer program product for designing combinatorial arrays
AU2001247627A1 (en) 2000-03-22 2001-10-03 3-Dimensional Pharmaceuticals, Inc. System, method, and computer program product for representing object relationships in a multidimensional space
US7139739B2 (en) 2000-04-03 2006-11-21 Johnson & Johnson Pharmaceutical Research & Development, L.L.C. Method, system, and computer program product for representing object relationships in a multidimensional space
WO2002061419A1 (en) 2001-01-29 2002-08-08 3-Dimensional Pharmaceuticals, Inc. Method, system, and computer program product for analyzing combinatorial libraries
US20030059811A1 (en) * 2001-06-14 2003-03-27 Hakim Djaballah Methods of screening for ligands of target molecules
EP1637885A1 (en) * 2004-09-16 2006-03-22 Vivalis Method of screening by using conformation sensitive peptides
GB0421285D0 (en) 2004-09-24 2004-10-27 Univ Nottingham Improvements in high content screening
US8158340B2 (en) 2005-12-28 2012-04-17 Corning Incorporated Methods for detecting conformational changes in bioentities
US10571471B2 (en) 2015-02-05 2020-02-25 Promega Corporation Luciferase-based thermal shift assays
GB202118173D0 (en) * 2021-12-15 2022-01-26 Medicines Discovery Catpult Ltd Protein stability assay

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5231755B2 (en) * 1973-01-08 1977-08-17
US5200504A (en) * 1990-10-02 1993-04-06 The Scripps Research Institute Metallopeptides having stabilized secondary structures
CA2115900A1 (en) * 1993-02-22 1994-08-23 Gerald W. Becker Pharmaceutical screens and antibodies

Also Published As

Publication number Publication date
WO1997020952A1 (en) 1997-06-12
JP2000502440A (en) 2000-02-29
EP0865502A4 (en) 2000-08-16
CA2236339A1 (en) 1997-06-12
EP0865502A1 (en) 1998-09-23

Similar Documents

Publication Publication Date Title
DE865502T1 (en) FLUORESCENT-BASED DETECTION METHOD FOR LIGANDS
EP1136822B1 (en) Method for identifying cell specific target structures
DE69824099T2 (en) ANALYTICAL PROCEDURE, TEST SET AND DEVICE
DE770876T1 (en) Screening method for ligand identification of target proteins
DE69828502T2 (en) Method for detecting biological molecules in biological samples by means of bacteriophages
DE19709348A1 (en) Automatic multi-epitope ligand mapping system
NZ510977A (en) Novel G protein-coupled receptor proteins, DNAs thereof and ligands to the same
DE69032847T2 (en) Hydrophobic nucleic acid probe
DE03808454T1 (en) PROCEDURE FOR PRESENTING THE EFFECT OF HER2 TARGETED THERAPY
Traut et al. Ribosomal protein L7/L12 cross-links to proteins in separate regions of the 50 S ribosomal subunit of Escherichia coli.
DE69919503T2 (en) METHOD FOR DETERMINING AN ANALYTE AND KIT THEREFOR
DE69924147T2 (en) FLOORESCENCE INTENSITY TEST FOR A PROTEIN OR PEPTIDE BINDING ON NUCLEIC ACIDS
DE1105508T1 (en) METHOD FOR PURIFYING PROTEINS AND BIOMOLECOLIC OR PROTEIN COMPLEXES
EP3165920B1 (en) Method for analysis of serum on anti-erythrocyte antibodies
DE69718624T2 (en) NON-COMPETITIVE IMMUNOASSAY WITH BLOCKING OF FREE BINDING POINTS ON THE SOLID PHASE
DE69931467T2 (en) NEW COMPLEXES CONTAINING NETWORKED AVIDIN, METHOD OF MANUFACTURING AND ANALYSIS METHOD USING THIS
EP1078262A1 (en) Indicator strip for the detection of antibodies in blood serum
WO2002073209A2 (en) Method for identifying immune reactive epitopes on proteins
DE2733380A1 (en) Immunological assay using carrier bound reactant - incubated with second reactant and then with labelled reactant specific for the second
Callaci et al. Assembly of Xenopus transcription factor III A-5S RNA complex
EP2020601A1 (en) Kit and device for creating chemo luminescent radiation
EP1441035B1 (en) Method of obtaining profiles of differential protein expression
DE69925736T2 (en) LUPUS ANTIKOAGULANT ANTIBODY AGAINST THE F1 REGION OF PROTHROMBIN
DE60209273T2 (en) Chip made of biological materials
DE2750521A1 (en) DIAGNOSTIC PROCEDURE