发明公开
- 专利标题: BIOLOGICAL ASSAY DETECTION METHOD
- 专利标题(中): 生物测定检测方法
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申请号: EP02768394.5申请日: 2002-08-02
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公开(公告)号: EP1442296A2公开(公告)日: 2004-08-04
- 发明人: INGLESE, James , FERRER, Marc , HAMILTON, Aaron, C.
- 申请人: Merck & Co., Inc.
- 申请人地址: 126 East Lincoln Avenue Rahway,New Jersey 07065-0907 US
- 专利权人: Merck & Co., Inc.
- 当前专利权人: Merck & Co., Inc.
- 当前专利权人地址: 126 East Lincoln Avenue Rahway,New Jersey 07065-0907 US
- 代理机构: Horgan, James Michael Frederic
- 优先权: US310599P 20010807
- 国际公布: WO2003014698 20030220
- 主分类号: G01N33/53
- IPC分类号: G01N33/53 ; G01N33/566 ; C12Q1/34 ; C12Q1/42
摘要:
The invention is a method for detecting a reaction product which signals the presence of a reaction product inducer such as an enzyme. The method enables the recognition of epitopes that form the basis of a detection strategy without the need for specific antibodies to the epitope. In the method, a directly or indirectly labeled modular domain and a biotinylated form of the cognate peptide ligand are used as the basis for a measurable interaction. The peptide ligand can be masked by modifications through, for example, phosphorylation of the Ser or Thr residue, or extension of the amino acid sequence beyond the C-terminal Val. Because the masked residues are critical to binding of the labeled modular domain, masking of at least one of the residues prevents binding. Upon treatment of the masked residue by the appropriate enzyme, (e.g., treatment of the phosphorylated residue with a phosphatase enzyme, or treatment of the extended residue with a protease enzyme, the peptide is converted to the original unmasked ligand that is capable of binding to the labeled modular domain and forming a measurable complex.
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