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WO2005038460A3 - Site-specific labeling of affinity tags in fusion proteins - Google Patents

Site-specific labeling of affinity tags in fusion proteins Download PDF

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Publication number
WO2005038460A3
WO2005038460A3 PCT/US2004/034460 US2004034460W WO2005038460A3 WO 2005038460 A3 WO2005038460 A3 WO 2005038460A3 US 2004034460 W US2004034460 W US 2004034460W WO 2005038460 A3 WO2005038460 A3 WO 2005038460A3
Authority
WO
WIPO (PCT)
Prior art keywords
affinity tag
integer
present
fusion proteins
fluorescent compounds
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/US2004/034460
Other languages
French (fr)
Other versions
WO2005038460A2 (en
Inventor
Kyle Gee
Courtenay Hart
Richard Haugland
Wayne Patton
Scott Whitney
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.)
Molecular Probes Inc
Original Assignee
Molecular Probes 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 Molecular Probes Inc filed Critical Molecular Probes Inc
Publication of WO2005038460A2 publication Critical patent/WO2005038460A2/en
Publication of WO2005038460A3 publication Critical patent/WO2005038460A3/en
Anticipated expiration legal-status Critical
Ceased 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/13—Labelling of peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biotechnology (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a polyhistidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6. The acetic acid groups interact directly with the positively charged histidine residues of the affinity tag to effectively label and detect a fusion protein containing such an affinity tag when present in an acidic or neutral environment.
PCT/US2004/034460 2003-10-14 2004-10-14 Site-specific labeling of affinity tags in fusion proteins Ceased WO2005038460A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US51125203P 2003-10-14 2003-10-14
US60/511,252 2003-10-14

Publications (2)

Publication Number Publication Date
WO2005038460A2 WO2005038460A2 (en) 2005-04-28
WO2005038460A3 true WO2005038460A3 (en) 2006-03-09

Family

ID=34465207

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2004/034460 Ceased WO2005038460A2 (en) 2003-10-14 2004-10-14 Site-specific labeling of affinity tags in fusion proteins

Country Status (1)

Country Link
WO (1) WO2005038460A2 (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5223162B2 (en) * 2006-02-28 2013-06-26 公立大学法人名古屋市立大学 Fluorescent dye for labeling peptide or protein, and labeling method using the fluorescent dye
US9535070B2 (en) * 2012-11-09 2017-01-03 Arizona Board Of Regents, A Body Corporate Of The State Of Arizona, Acting For And On Behalf Of Arizona State University High throughput detection of fusion proteins
CN113943372A (en) * 2021-11-09 2022-01-18 南京工业大学 A method for improving the stability of immobilized protein by using trivalent cobalt ions

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002012195A1 (en) * 2000-08-04 2002-02-14 Molecular Probes, Inc. Derivatives of 1,2-dihydro-7-hydroxyquinolines containing fused rings

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002012195A1 (en) * 2000-08-04 2002-02-14 Molecular Probes, Inc. Derivatives of 1,2-dihydro-7-hydroxyquinolines containing fused rings

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
KAPANIDIS A N ET AL: "Site-specific incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (Ni(2+):nitrilotriacetic Acid (n)-fluorochrome conjugates", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, AMERICAN CHEMICAL SOCIETY, WASHINGTON, DC, US, vol. 123, no. 48, 5 December 2001 (2001-12-05), pages 12123 - 12125, XP002257748, ISSN: 0002-7863 *
STORA T ET AL: "METAL ION TRACE DETECTION BY A CHELATOR-MODIFIED GOLD ELECTRODE: A COMPARISON OF SURFACE TO BULK AFFINITY", LANGMUIR, AMERICAN CHEMICAL SOCIETY, NEW YORK, NY, US, vol. 13, no. 20, 1 October 1997 (1997-10-01), pages 5212 - 5214, XP001040532, ISSN: 0743-7463 *

Also Published As

Publication number Publication date
WO2005038460A2 (en) 2005-04-28

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