Abstract: The invention discloses newly-discovered phosphorylation sites in human IRS-1 and IRS-2, serine 1101 (Ser1101) and serine 1149 (Ser1149) respectively, and provides antibodies, both polyclonal and monoclonal, that selectively bind to IRS-1 and/or IRS-2 when phosphorylated at these respective sites, but do not bind to IRS-1 and/or IRS-2 when not phosphorylated at these respective sites. The sites are relevant to insulin-resistance in type 2 diabetes. Also provided are methods for determining the phosphorylation of IRS-1/2 or activity of PKC theta in a biological sample, by using a detectable reagent, such as the disclosed antibodies, that binds to IRS-1/2 only when phosphorylated at Ser1101/Ser1149. Kits comprising the phosphor-IRS-1/2 (Ser1101/1149) antibodies of the invention are also provided.
Abstract: In accordance with the invention, novel gene deletions and translocations involving chromosome 2 resulting in fusion proteins combining part of Anaplastic Lymphoma Kinase (ALK) kinase with part of a secondary protein have now been identified in human solid tumors, e.g. non-small cell lung carcinoma (NSCLC). Secondary proteins include Echinoderm Microtubule-Associated Protein-Like 4 (EML-4) and TRK-Fusion Gene (TFG). The EML4-ALK fusion protein, which retains ALK tyrosine kinase activity, was confirmed to drive the proliferation and survival of NSCLC characterized by this mutation. The invention therefore provides, in part, isolated polynucleotides and vectors encoding the disclosed mutant ALK kinase polypeptides, probes for detecting it, isolated mutant polypeptides, recombinant polypeptides, and reagents for detecting the fusion and truncated polypeptides.
Type:
Grant
Filed:
April 13, 2007
Date of Patent:
April 20, 2010
Assignee:
Cell Signaling Technology, Inc.
Inventors:
Klarisa Rikova, Herbert Haack, Laura Sullivan, Ailan Guo, Anthony Possemato, Joan MacNeill, Ting-Lei Gu, Jian Yu
Abstract: The invention discloses 443 novel phosphorylation sites identified in leukemia, peptides (including AQUA peptides) comprising a phosphorylation site of the invention, antibodies specifically bind to a novel phosphorylation site of the invention, and diagnostic and therapeutic uses of the above.
Type:
Application
Filed:
September 18, 2007
Publication date:
December 31, 2009
Applicant:
Cell Signaling Technology , Inc
Inventors:
Peter Hornbeck, Albrecht Moritz, Valerie Goss, Kimberly Lee, Ting-Lie Gu
Abstract: The present invention is related to a method for producing motif-specific, context-independent antibodies which are specific to at least one modified amino acid residue in the context of variable surrounding amino acid or peptide sequences. The method is particularly useful in producing antibodies which recognize phosphorylated serine, threonine, and tyrosine, or acetylated lysine, as well as other modified amino acids-containing motifs of one or more amino acids.
Abstract: A method is provided for producing motif-specific, context-independent antibodies which recognize a plurality of peptides or proteins within a genome that contain the same motif. The method includes the step of immunizing a host with a degenerate peptide library antigen featuring (i) a fixed target motif containing one or more invariant amino acids including at least one modified amino acid, and (ii) a plurality of degenerate amino adds flanking the motif. Motif-specific, context-independent antibodies produced by the disclosed method are also provided. The method encompasses motifs consisting of a single modified amino acid, as well as short motifs comprising multiple invariant amino acids including one or more modified amino acids, such as all or part of kinase consensus substrate motifs, protein-protein binding motifs, or other cell signaling motifs. Methods of using the antibodies, e.g. for genome-wide profiling, are also provided.
Abstract: The invention provides methods for isolating a modified peptide from a complex mixture of peptides, the method comprising the steps of: (a) obtaining a proteinaceous preparation from an organism, wherein the preparation comprises modified peptides from two or more different proteins; (b) contacting the preparation with at least one immobilized modification-specific antibody; and (c) isolating at least one modified peptide specifically bound by the immobilized modification-specific antibody in step (b). The method may further comprise the step of (d) characterizing the modified peptide isolated in step (c) by mass spectrometry (MS), tandem mass spectrometry (MS—MS), and/or MS3 analysis, or the step of (e) utilizing a search program to substantially match the spectra obtained for the modified peptide during the characterization of step (d) with the spectra for a known peptide sequence, thereby identifying the parent protein(s) of the modified peptide.
Type:
Grant
Filed:
June 19, 2002
Date of Patent:
April 3, 2007
Assignee:
Cell Signaling Technology, Inc.
Inventors:
John Rush, Hui Zhang, Xiangming Zha, Michael J. Comb, Yi Tan
Abstract: The invention discloses two newly-discovered Flt3 phosphorylation sites, tyrosine 589 (Tyr589) and tyrosine 591 (Tyr591) in the intracellular domain, and provides antibodies, both polyclonal and monoclonal, that selectively bind to Flt3 when phosphorylated at these novel sites. Also provided are assays utilizing these reagents, including methods for determining the phosphorylation of Flt3 in a biological sample, selecting a patient suitable for Flt3 inhibitor therapy, profiling Flt3 activation in a test tissue, and identifying a compound that modulates phosphorylation of Flt3 in a test tissue, by using a detectable reagent, such as the disclosed antibodies, that binds to Flt3 when phosphorylated at Tyr589 or Tyr591. The sample or test tissue may be taken from a subject suspected of having cancer, such as acute myelogenous leukemia (AML).
Type:
Grant
Filed:
February 18, 2003
Date of Patent:
February 27, 2007
Assignee:
Cell Signaling Technology, Inc.
Inventors:
Michael J. Comb, Randall K. Wetzel, Jiong Wu, Katherine Crosby
Abstract: The invention provides monoclonal antibodies that bind the estrogen receptor ? (ER ?) when phosphorylated at serine 118 (Ser118) in the N-terminal domain, but do not bind to ER ? when not phosphorylated at this site. Also provided are methods for determining the phosphorylation of ER ? in a biological sample, profiling ER ? activation in a test tissue, and identifying a compound that modulates phosphorylation of ER ? in a test tissue, by using the disclosed monoclonal antibodies. The sample or test tissue may be taken from a subject suspected of having cancer, such as breast cancer. Kits comprising the phospho-ER ? (Ser118) monoclonal antibodies of the invention are also provided.
Type:
Grant
Filed:
August 2, 2002
Date of Patent:
September 12, 2006
Assignee:
Cell Signalling Technology, Inc.
Inventors:
Bradley L. Smith, Katherine Crosby, Jiong Wu
Abstract: A class of motif-specific, context-independent antibodies that bind conserved signal transduction motifs, such as kinase consensus substrate motifs and protein-protein binding motifs, containing one or more modified amino acids, such as a phosphorylated amino acid, is provided. The antibodies bind a plurality of peptides or proteins that contain the modified motif. Context-independent antibodies specific for single modified residues, such as phosphothreonine, are also provided. Methods for producing and using such antibodies are provided.
Abstract: The invention discloses newly-discovered phosphorylation sites in human IRS-1 and IRS-2, serine 1101 (Ser1101) and serine 1149 (Ser1149) respectively, and provides antibodies, both polyclonal and monoclonal, that selectively bind to IRS-1 and/or IRS-2 when phosphorylated at these respective sites, but do not bind to IRS-1 and/or IRS-2 when not phosphorylated at these respective sites. The sites are relevant to insulin-resistance in type 2 diabetes. Also provided are methods for determining the phosphorylation of IRS-1/2 or activity of PKC theta in a biological sample, by using a detectable reagent, such as the disclosed antibodies, that binds to IRS-1/2 only when phosphorylated at Ser1101/Ser1149. Kits comprising the phosphor-IRS-1/2 (Ser1101/1149) antibodies of the invention are also provided.
Abstract: The invention provides novel reagents and methods for detecting BCR-ABL or c-Abl kinase activity, and/or Abl signaling pathway activation in a cell or tissue, and discloses novel biomarkers relevant to Abl-mediated disease progression and therapeutic responsiveness, and provides predictive and detection methods based on the same. Phosphorylated BCR-ABL (Tyr245) and/or c-Abl (Tyr245), BCR-ABL (Tyr735) and/or c-Abl (Tyr735), Bcr (Tyr177), CRKL (Tyr207), Gab1 (Tyr627), PYK2 (Tyr402), Tyk2 (Tyr1054/1055), SHP2 (Tyr580), ERK1/2 (Thr202/Tyr204) and MEK1/2 (Ser217/221) have now been identified as relevant biomarkers of c-Abl pathway-mediated disease, and phospho-specific antibodies to these targets are provided. Kits for carrying out the methods of the invention are also provided.
Type:
Application
Filed:
April 7, 2003
Publication date:
October 9, 2003
Applicant:
Cell Signaling Technology, Inc.
Inventors:
Katherine Crosby, Bradley L. Smith, Valerie L. Goss
Abstract: The invention provides a method for producing antibodies that selectively recognize short, modified amino acid motifs substantially independent of the surrounding amino acid context in which the motif occurs. A novel class of motif-specific, context-independent antibodies is also provided. The invention encompasses modified motifs consisting of single modified amino acids, for example phosphotyrosine or acetylated lysine, as well other modified motifs of multiple amino acids, such as kinase consensus substrate motifs and protein-protein binding motifs relevant to cell signal transduction. Also provided are methods of profiling large and diverse protein populations on a genome-wide basis by utilizing the antibodies of the invention, and methods for the positive identification of cellular phosphoproteins using one or more motif-specific, context-independent antibodies of the invention coupled with protein database searching.
Abstract: The present invention is related to a method for producing motif-specific, context-independent antibodies which are specific to at least one modified fixed amino acid residue in the context of variable surrounding amino acid or peptide sequences. The method is particularly useful in producing antibodies which recognize phosphorylated serine, threonine, and tyrosine, or acetylated lysine, as well as other modified amino acid-containing motifs of one or more amino acids.