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    MONITORING PROTEIN TRAFFICKING USING BETA-GALACTOSIDASE REPORTER FRAGMENT COMPLEMENTATION
    1.
    发明公开
    MONITORING PROTEIN TRAFFICKING USING BETA-GALACTOSIDASE REPORTER FRAGMENT COMPLEMENTATION 有权
    监测β-半乳糖苷记者-FRAGMENTKOMPLEMENTIERUNG的蛋白质运动

    公开(公告)号：EP2724158A1

    公开(公告)日：2014-04-30

    申请号：EP12802981.6

    申请日：2012-06-22

    申请人： DiscoveRx Corporation

    发明人： WEHRMAN, Thomas, S. ,  BASSONI, Daniel ,  RAAB, William

    IPC分类号： G01N33/53 ,  C12Q1/02

    CPC分类号： C12Q1/34 ,  C07K2319/61 ,  G01N33/5076

    摘要： Methods and materials are disclosed for use in an enzyme fragment complementation assay using complementary fragments of β-galactosidase to study the trafficking of proteins in a cell. Compounds that bind to a target peptide have been found to affect protein folding and therefore trafficking. β-Galactosidase fragments, an enzyme donor (ED) and an enzyme acceptor (EA), are fused to a target peptide and to an intracellular compartment protein, wherein the compartment is involved in intracellular trafficking. Contacting the cell with a compound that binds to the target peptide results in enhanced movement of the protein through the cellular trafficking pathway comprised of the endoplasmic reticulum, Golgi apparatus, the plasma membrane, endosomes, etc. Using this approach, compounds that bind to a target peptide and alter its ability to traffic through the normal cellular pathway can be readily detected.

    HOMOGENOUS THERMAL SHIFT LIGAND BINDING ASSAY
    3.
    发明公开
    HOMOGENOUS THERMAL SHIFT LIGAND BINDING ASSAY 审中-公开
    同源热转移配体结合测定

    公开(公告)号：EP3183580A1

    公开(公告)日：2017-06-28

    申请号：EP15834042.2

    申请日：2015-08-18

    申请人： DiscoveRx Corporation

    发明人： TREIBER, Daniel K. ,  MENICHELLI, Elena

    IPC分类号： G01N33/58 ,  G01N33/68

    CPC分类号： G01N33/581 ,  G01N33/542 ,  G01N2333/938 ,  G01N2500/04

    摘要： Disclosed are methods for detecting and quantitatively measuring a binding property of a compound to a target macromolecule, wherein the target macromolecule is subject to denaturation and is linked to a labeling peptide, such as a short enzyme fragment. The method uses a fluid mixture comprising (i) a chimeric molecule comprising a target macromolecule linked to the labeling peptide and (ii) a compound being measured for binding to the target macromolecule, wherein said target macromolecule is subject to denaturation. After allowing for binding of the compound (e.g. a small molecule inhibitor of the target macromolecule), one detects a signal from the labeling peptide, such as by enzyme fragment complementation. This signal indicates a differential between denatured and non-denatured target macromolecules and thereby indicates a differential between target macromolecules not bound to the compound and target macromolecules bound to the compound, respectively.

    摘要翻译： 公开了用于检测和定量测量化合物与靶大分子的结合性质的方法，其中靶大分子经历变性并连接至标记肽，如短酶片段。 所述方法使用流体混合物，所述流体混合物包含（i）包含与标记肽连接的靶大分子的嵌合分子和（ii）测量与靶大分子结合的化合物，其中所述靶大分子经历变性。 在允许化合物（例如目标大分子的小分子抑制剂）结合之后，检测来自标记肽的信号，例如通过酶片段互补。 该信号表明变性和未变性靶标大分子之间的差异，并且由此分别指示未与化合物结合的靶标大分子和与化合物结合的靶标大分子之间的差异。

    RECEPTOR TYROSINE KINASE ASSAYS
    9.
    发明公开
    RECEPTOR TYROSINE KINASE ASSAYS 有权
    受体测定

    公开(公告)号：EP2313517A1

    公开(公告)日：2011-04-27

    申请号：EP09822362.1

    申请日：2009-08-06

    申请人： DiscoveRx Corporation

    发明人： FENG, Wei ,  RAAB, William ,  ACHACOSO, Philip ,  WEHRMAN, Thomas ,  OLSON, Keith, R.

    IPC分类号： C12Q1/00

    CPC分类号： G01N33/566 ,  C12Q1/34 ,  C12Q1/485

    摘要： Methods for detecting phosphorylation of receptor tyrosine kinases ("RTKs") upon activation are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of β-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of β-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a β- galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.