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公开(公告)号：US08647887B2
公开(公告)日：2014-02-11
申请号：US13141661
申请日：2010-01-28
IPC分类号： G01N33/567 , C07K14/705 , C07K16/00
CPC分类号： G01N33/542 , G01N2333/726
摘要： The present invention relates to methods and polypeptides for detecting a compound in a sample. In particular, the present invention relates to the use of a cell-free composition comprising at least one G protein coupled receptor embedded in a lipid Mayer which when expressed in a cell the N-terminus of the G protein coupled receptor, or subunits thereof, is outside the cell and the C-terminus is inside the cell, and which is capable of binding the compound. Optionally, the composition also comprises at least one accessory molecule that directly or indirectly binds an intracellular loop and/or the C-terminus of the G protein coupled receptor. The G protein coupled receptor, and/or accessory molecule when present, in combination comprise a bioluminescent protein and an acceptor molecule, which enables bioluminescent resonance energy transfer (BRET) to be used to detect the compound binding the receptor.
摘要翻译： 本发明涉及用于检测样品中化合物的方法和多肽。 特别地，本发明涉及包含至少一种嵌入脂质Mayer中的G蛋白偶联受体的无细胞组合物的用途，当在细胞中表达G蛋白偶联受体的N末端或其亚基时， 在细胞外，C末端位于细胞内，并且能够结合化合物。 任选地，组合物还包含直接或间接结合G蛋白偶联受体的细胞内环和/或C末端的至少一个辅助分子。 当存在时，G蛋白偶联受体和/或辅助分子组合包含生物发光蛋白和受体分子，其使得能够使用生物发光共振能量转移（BRET）来检测结合受体的化合物。
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公开(公告)号：US20120077210A1
公开(公告)日：2012-03-29
申请号：US13141661
申请日：2010-01-28
IPC分类号： C12Q1/66
CPC分类号： G01N33/542 , G01N2333/726
摘要： The present invention relates to methods and polypeptides for detecting a compound in a sample. In particular, the present invention relates to the use of a cell-free composition comprising at least one G protein coupled receptor embedded in a lipid Mayer which when expressed in a cell the N-terminus of the G protein coupled receptor, or subunits thereof, is outside the cell and the C-terminus is inside the cell, and which is capable of binding the compound. Optionally, the composition also comprises at least one accessory molecule that directly or indirectly binds an intracellular loop and/or the C-terminus of the G protein coupled receptor. The G protein coupled receptor, and/or accessory molecule when present, in combination comprise a bioluminescent protein and an acceptor molecule, which enables bioluminescent resonance energy transfer (BRET) to be used to detect the compound binding the receptor.
摘要翻译： 本发明涉及用于检测样品中化合物的方法和多肽。 特别地，本发明涉及包含至少一种嵌入脂质Mayer中的G蛋白偶联受体的无细胞组合物的用途，当在细胞中表达G蛋白偶联受体的N末端或其亚基时， 在细胞外，C末端位于细胞内，并且能够结合化合物。 任选地，组合物还包含直接或间接结合G蛋白偶联受体的细胞内环和/或C末端的至少一个辅助分子。 当存在时，G蛋白偶联受体和/或辅助分子组合包含生物发光蛋白和受体分子，其使得能够使用生物发光共振能量转移（BRET）来检测结合受体的化合物。
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