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WO2006113050A4 - Reagents for the detection of protein phosphorylation in carcinoma signaling pathways - Google Patents

Reagents for the detection of protein phosphorylation in carcinoma signaling pathways Download PDF

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Publication number
WO2006113050A4
WO2006113050A4 PCT/US2006/010868 US2006010868W WO2006113050A4 WO 2006113050 A4 WO2006113050 A4 WO 2006113050A4 US 2006010868 W US2006010868 W US 2006010868W WO 2006113050 A4 WO2006113050 A4 WO 2006113050A4
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WO
WIPO (PCT)
Prior art keywords
phosphorylated
antibody
column
tyrosine
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.)
Ceased
Application number
PCT/US2006/010868
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French (fr)
Other versions
WO2006113050A8 (en
WO2006113050A3 (en
WO2006113050A2 (en
Inventor
Ailan Guo
Klarisa Rikova
Albrecht Moritz
Yu Li
Charles Farnsworth
Kimberly Lee
Roberto Polakiewicz
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.)
Cell Signaling Technology Inc
Original Assignee
Cell Signaling Technology Inc
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Filing date
Publication date
Application filed by Cell Signaling Technology Inc filed Critical Cell Signaling Technology Inc
Priority to EP06739581A priority Critical patent/EP1872134A4/en
Publication of WO2006113050A2 publication Critical patent/WO2006113050A2/en
Anticipated expiration legal-status Critical
Publication of WO2006113050A8 publication Critical patent/WO2006113050A8/en
Publication of WO2006113050A3 publication Critical patent/WO2006113050A3/en
Publication of WO2006113050A4 publication Critical patent/WO2006113050A4/en
Priority to US12/985,879 priority patent/US20110105732A1/en
Ceased legal-status Critical Current

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Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57496Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving intracellular compounds
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4748Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/44Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Cell Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Urology & Nephrology (AREA)
  • Biophysics (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Food Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention discloses (214) novel phosphorylation sites identified in signal transduction proteins and pathways underlying human carcinoma, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Adaptor/Scaffold proteins, Cytoskeleton proteins, GTP Signaling proteins, Kinases, Metabolism proteins, Phosphatases/Phospho- diesterases/ Proteases, Receptor proteins, RNA Processing proteins, Transcription proteins, Translation proteins, Transporter proteins, and Ubitquitin proteins, as well as other protein types.

Claims

82
AMENDED CLAIMS [Received by the International Bureau on 22 DEC 2008 (22.12.2008)]
49. A method selected from the group consisting of:
5 (a) a method for detecting a human carcinoma-related signaling protein selected from Rows 139, 123, 93, 97, 45, 94, 122 and 124 in Column A of Table 1, wherein said human carcinoma-related signaling protein is phosphorylated at the tyrosine listed in corresponding Column D of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E of Table 1 (SEQ ID NOs: 138,0 122, 92, 9644, 93, 121 and 123), comprising the step of adding an isolated phosphorylation-specific antibody according to claim 14, to a sample comprising said human carcinoma-related signaling protein under conditions that permit the binding of said antibody to said human carcinoma-related signaling protein, and detecting bound antibody; 5 (b) a method for quantifying the amount of a human carcinoma-related signaling protein listed in Rows 139, 123, 93, 97, 45, 94, 122 and 124 in Column A of Table 1 that is phosphorylated at the corresponding tyrosine listed in Column D of Table 1 , comprised within the phosphorylatable peptide sequence listed in corresponding Column E of Table 1 (SEQ ID NOs: 138, 122, 92, 96 44, 93, 121 and 123), in a 0 sample using a heavy-isotope labeled peptide (AQUA ™ peptide), said labeled peptide comprising a phosphorylated tyrosine at said corresponding tyrosine listed Column D of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E of Table 1 (SEQ ID NOs: 138, 122, 92, 9644, 93, 121 and 123) as an internal standard; and 5 (c) a method comprising step (a) followed by step (b).
50. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding PDGFRa only when phosphorylated at Y849, comprised within the phosphorylatable peptide sequence listed in Column E, Row 139, of Table 1 (SEQ ID NO: 138), wherein said antibody does not bind said 0 protein when not phosphorylated at said tyrosine.
51. The method of claim 49, wherein said isolated phosphorylation-specific 83 antibody is capable of specifically binding PDGFRa only when not phosphorylated at Y849, comprised within the phosphorylatable peptide sequence listed in Column E, Row 139, of Table 1 (SEQ ID NO: 138), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
52. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding LYN only when phosphorylated at Y 193, comprised within the phosphorylatable peptide sequence listed in Column E, Row 123, of Table 1 (SEQ ID NO: 122), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.
53. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding LYN only when not phosphorylated at Yl 93, comprised within the phosphorylatable peptide sequence listed in Column E, Row 123, of Table 1 (SEQ ID NO: 122), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
54. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding Cdk2, only when phosphorylated at Y 19, comprised within the phosphoiylatable peptide sequence listed in Column E, Row 93, of Table 1 (SEQ ID NO: 92), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.
55. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding Cdk2 only when not phosphorylated at Y 19, comprised within the phosphoiylatable peptide sequence listed in Column E, Row 93, of Table 1 (SEQ ID NO: 92), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
56. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding DYRK4 only when phosphorylated at Y264, comprised within the phosphorylatable peptide sequence listed in Column E, Row 97, of Table 1 (SEQ ID NO: 96), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine. 84
57. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding DYK4 only when not phosphorylated at Y264, comprised within the phosphorylatable peptide sequence listed in Column E, Row 97, of Table 1 (SEQ ID NO: 96), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
58. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding actin, beta only when phosphorylated at Y 169, comprised within the phosphorylatable peptide sequence listed in Column E, Row 45, of Table 1 (SEQ ID NO: 44), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.
59. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding actin, beta only when not phosphorylated at Y 169, comprised within the phosphorylatable peptide sequence listed in Column E, Row 45, of Table 1 (SEQ ID NO: 44), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
60. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding Cdk2 only when phosphorylated at Yl 5, comprised within the phosphorylatable peptide sequence listed in Column E, Row 94, of Table I (SEQ ID NO: 93), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.
61. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding Cdk2 only when not phosphorylated at Yl 5, comprised within the phosphorylatable peptide sequence listed in Column E, Row 94, of Table 1 (SEQ ID NO: 93), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
62. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding Fyn only when phosphorylated at Y439, comprised within the phosphorylatable peptide sequence listed in Column E, Row 122, of Table 1 (SEQ ID NO: 121), wherein said antibody does not bind said protein 85 when not phosphorylated at said tyrosine.
63. The method of claim 49, wherein said isolated phosphorylation-specifϊc antibody is capable of specifically binding Fyn only when not phosphorylated at Y439, comprised within the phosphorylatable peptide sequence listed in Column E, Row 122, of Table 1 (SEQ ID NO: 121), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
64. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding AxI only when phosphorylated at Y696, comprised within the phosphorylatable peptide sequence listed in Column E, Row 124, of Table 1 (SEQ ID NO: 123), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.
65. The method of claim 49, wherein said isolated phosphorylation-specific antibody is capable of specifically binding AxI only when not phosphorylated at Y696, comprised within the phosphorylatable peptide sequence listed in Column E, Row 124, of Table 1 (SEQ ID NO: 123), wherein said antibody does not bind said protein when phosphorylated at said tyrosine.
PCT/US2006/010868 2005-04-12 2006-03-24 Reagents for the detection of protein phosphorylation in carcinoma signaling pathways Ceased WO2006113050A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP06739581A EP1872134A4 (en) 2005-04-12 2006-03-24 REAGENTS FOR DETECTION OF PHOSPHORYLATION OF PROTEINS IN THE CARCINOMA SIGNALING PATHWAY
US12/985,879 US20110105732A1 (en) 2005-04-12 2011-01-06 Reagents for the Detection of Protein Phosphorylation in Carcinoma Signaling Pathways

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US67044705P 2005-04-12 2005-04-12
US60/670,447 2005-04-12

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
US67044705P Continuation 2005-04-12 2005-04-12

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US11/974,480 Continuation US20090099340A1 (en) 2005-04-12 2007-10-12 Reagents for the detection of protein phosphorylation in carcinoma signaling pathways

Publications (4)

Publication Number Publication Date
WO2006113050A2 WO2006113050A2 (en) 2006-10-26
WO2006113050A8 WO2006113050A8 (en) 2007-12-27
WO2006113050A3 WO2006113050A3 (en) 2008-12-24
WO2006113050A4 true WO2006113050A4 (en) 2009-03-26

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EP (1) EP1872134A4 (en)
WO (1) WO2006113050A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8242083B2 (en) * 2008-06-23 2012-08-14 Perkinelmer Health Sciences, Inc. Kinase substrates
CN111116734A (en) * 2019-12-23 2020-05-08 维塔恩(广州)医药有限公司 Tumor-associated gene c-kit mutation-associated antigen short peptide and application thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0640102B1 (en) * 1992-04-10 2006-08-09 Dana-Farber Cancer Institute, Inc. Activation-state-specific phosphoprotein immunodetection
WO2002081638A2 (en) * 2001-04-06 2002-10-17 Origene Technologies, Inc Prostate cancer expression profiles
US7335467B2 (en) * 2001-05-15 2008-02-26 Ludwig Institute For Cancer Research Breast cancer antigens

Also Published As

Publication number Publication date
WO2006113050A8 (en) 2007-12-27
EP1872134A2 (en) 2008-01-02
WO2006113050A3 (en) 2008-12-24
WO2006113050A2 (en) 2006-10-26
EP1872134A4 (en) 2010-09-01

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