Reagents for the detection of protein phosphorylation in anaplastic large cell lymphoma signaling pathways

• US 7,973,134 B2
• Filed: 08/11/2006
• Issued: 07/05/2011
• Est. Priority Date: 07/07/2004
• Status: Active Grant

• Current Assignee: Cell Signaling Technology Incorporated
• Original Assignee: Cell Signaling Technology Incorporated
• Inventors: Lee, Kimberly, Moritz, Albrecht, Rush, John, Polakiewicz, Roberto
• Primary Examiner(s): Goddard; Laura B
• Application Number:
  US11/503,096
• Publication Number:
  US 20070072235A1
• Time in Patent Office: 1,789 Days
• Field of Search: None
• US Class Current: 530/387.1
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: G01N 33/6842 Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins

Abstract

The invention discloses 211 novel phosphorylation sites identified in signal transduction proteins and pathways underlying Anaplastic Large Cell Lymphoma (ALCL) involving the ALK-NPM translocation/fusion, 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: Protein Kinases (including Receptor Tyrosine Kinases), Adaptor/Scaffold Proteins, Cellular Metabolism or Miscellaneous Enzymes, Oxidoreductases, Transcription Factors, Cytoskeletal Proteins, Translation Initiation Complexes, RNA Binding Proteins, Proteases, Acetyltransferases, G protein regulators/GTPases, Helicases, Apoptosis/Cell Cycle Regulation proteins, and Hydrolases.

Claims

1. An isolated phosphorylation site-specific antibody that specifically binds a human Anaplastic Large Cell Lymphoma (ALCL)-related signaling protein, wherein said signaling protein is prohibitin D, only when said signaling protein is phosphorylated at the tyrosine at position 128, comprised within the phosphorylatable peptide sequence listed in SEQ ID NO:

40, wherein said antibody does not bind said signaling protein when not phosphorylated at said tyrosine.

2. An immortalized cell line producing the antibody of claim 1.

3. The cell line of claim 2, wherein said immortalized cell line is a rabbit hybridoma or a mouse hybridoma.