发明申请
- 专利标题: Biological assay detection method
- 专利标题(中): 生物检测方法
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申请号: US10484859申请日: 2002-08-02
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公开(公告)号: US20050164311A1公开(公告)日: 2005-07-28
- 发明人: James Inglese , Marc Ferrer , Aaron Hamilton
- 申请人: James Inglese , Marc Ferrer , Aaron Hamilton
- 国际申请: PCT/US02/24469 WO 20020802
- 主分类号: G01N33/531
- IPC分类号: G01N33/531 ; C07K5/10 ; C07K7/06 ; C07K7/08 ; C12Q1/34 ; C12Q1/37 ; C12Q1/42 ; C12Q1/48 ; C12Q1/527 ; G01N33/53 ; G01N33/542 ; G01N33/573
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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