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    METHODS FOR THE IDENTIFICATION OF LRRK2 INTERACTING MOLECULES AND FOR THE PURIFICATION OF LRRK2
    4.
    发明申请
    METHODS FOR THE IDENTIFICATION OF LRRK2 INTERACTING MOLECULES AND FOR THE PURIFICATION OF LRRK2 失效
    LRRK2相互作用分子鉴定方法及LRRK2纯化方法

    公开(公告)号：US20090220992A1

    公开(公告)日：2009-09-03

    申请号：US12282851

    申请日：2007-03-13

    申请人： Gerard Drewes ,  Carsten Hopf ,  Valerie Reader

    发明人： Gerard Drewes ,  Carsten Hopf ,  Valerie Reader

    IPC分类号： G01N33/573 ,  C07D403/04 ,  C12N9/12

    CPC分类号： C12Q1/485 ,  G01N2800/2835

    摘要： The invention provides in a first aspect a method for the identification of an LRRK2 interacting compound, comprising the steps of providing a protein preparation containing LRRK2, contacting the protein preparation with indol ligand 91 immobilized on a solid support under conditions allowing the formation of an indol ligand 91-LRRK2 complex, incubating the indol ligand 91-LRRK2 complex with a given compound, and determining whether the compound is able to separate LRRK2 from the immobilized indol ligand 91. Furthermore, the invention relates to a method for the identification of an LRRK2 interacting compound, comprising the steps of providing a protein preparation containing LRRK2, contacting the protein preparation with indol ligand 91 immobilized on a solid support and with a given compound under conditions allowing the formation of an indol ligand 91-LRRK2 complex, and detecting the indol ligand 91-LRRK2 complex. Additionally, the invention provides a method for the identification of an LRRK2 interacting compound, comprising the steps of providing two aliquots of a protein preparation containing LRRK2, contacting one aliquot with indol ligand 91 immobilized on a solid support under conditions allowing the formation of an indol ligand 91-LRRK2 complex, contacting the other aliquot with indol ligand 91 immobilized on a solid support and with a given compound under conditions allowing the formation of an indol ligand 91-LRRK2 complex, and determining the amount of indol ligand 91-LRRK2 complex. Furthermore, the invention relates to a method for the purification of LRRK2, comprising the steps of providing a protein preparation containing LRRK2, contacting the protein preparation with indol ligand 91 immobilized on a solid support under conditions allowing the formation of an indol ligand 91-LRRK2 complex, and separating LRRK2 from the immobilized indol ligand 91.

    摘要翻译： 本发明在第一方面提供了一种用于鉴定LRRK2相互作用化合物的方法，包括以下步骤：提供含有LRRK2的蛋白质制剂，使所述蛋白质制剂与固定在固体支持物上的吲哚配体91接触，所述条件允许形成吲哚 配体91-LRRK2复合物，将吲哚配体91-LRRK2复合物与给定化合物一起温育，并测定化合物是否能够从固定的吲哚配体91分离LRRK2。此外，本发明涉及用于鉴定LRRK2的方法 包括以下步骤：提供含有LRRK2的蛋白质制剂，使蛋白质制备物与固定在固体载体上的吲哚配体91与允许形成吲哚配体91-LRRK2复合物的条件下的给定化合物接触，并检测吲哚 配体91-LRRK2复合物。 另外，本发明提供了一种用于鉴定LRRK2相互作用化合物的方法，包括以下步骤：提供含有LRRK2的蛋白质制剂的两个等分试样，将一个等分试样与固定在固体支持物上的吲哚配体91接触，形成吲哚 配体91-LRRK2复合物，使其它等分试样与固定在固体载体上的吲哚配体91与给定化合物在允许形成吲哚配体91-LRRK2复合物的条件下接触，并测定吲哚配体91-LRRK2复合物的量。 此外，本发明涉及LRRK2的纯化方法，包括以下步骤：提供含有LRRK2的蛋白质制剂，使蛋白质制剂与固定在固体支持物上的吲哚配体91接触，形成吲哚配体91-LRRK2 复合物，并将LRRK2与固定的吲哚配体91分离。

    IN VIVO METHOD FOR THE EVALUATION OF A COMPOUND-TARGET INTERACTION
    5.
    发明申请
    IN VIVO METHOD FOR THE EVALUATION OF A COMPOUND-TARGET INTERACTION 审中-公开
    用于评估化合物 - 目标相互作用的体内方法

    公开(公告)号：US20130210030A1

    公开(公告)日：2013-08-15

    申请号：US13809811

    申请日：2011-06-20

    申请人： Gerard Drewes ,  Carsten Hopf ,  Gitte Neubauer ,  Ulrich Kruse

    发明人： Gerard Drewes ,  Carsten Hopf ,  Gitte Neubauer ,  Ulrich Kruse

    IPC分类号： G01N33/573

    CPC分类号： G01N33/573 ,  G01N33/5088 ,  G01N2333/91215 ,  G01N2500/00

    摘要： The invention relates to a method for evaluation of a compound-target interaction in vivo, comprising the steps of administering the compound to an animal, taking a body fluid sample of the animal, determining the concentration of the compound in the body fluid sample, taking a cellular sample of the animal containing the target, preparing a protein preparation of said cellular sample, providing an immobilised ligand capable of binding to the target, contacting the protein preparation with the immobilised ligand under conditions allowing the formation of a complex between the immobilised ligand and the target, determining the amount of complexes formed in step, and correlating the amount of complexes with the concentration of the compound in the body fluid sample.

    摘要翻译： 本发明涉及一种用于评价体内化合物 - 靶相互作用的方法，包括以下步骤：向动物施用化合物，服用动物的体液样品，测定体液样品中化合物的浓度，取 制备含有靶标的动物的细胞样品，制备所述细胞样品的蛋白质制剂，提供能够结合靶标的固定配体，使蛋白质制剂与固定的配体在允许形成固定化配体之间的复合物的条件下接触 和靶标，测定步骤中形成的复合物的量，并将复合物的量与体液样品中化合物的浓度相关联。

    METHODS FOR THE IDENTIFICATION OF ZAP-70 INTERACTING MOLECULES AND FOR THE PURIFICATION OF ZAP-70
    8.
    发明申请
    METHODS FOR THE IDENTIFICATION OF ZAP-70 INTERACTING MOLECULES AND FOR THE PURIFICATION OF ZAP-70 审中-公开

    公开(公告)号：US20120178180A1

    公开(公告)日：2012-07-12

    申请号：US12302711

    申请日：2007-06-01

    申请人： Ulrich Kruse ,  Nigel Ramsden ,  Gerard Drewes ,  Dirk Eberhard

    发明人： Ulrich Kruse ,  Nigel Ramsden ,  Gerard Drewes ,  Dirk Eberhard

    IPC分类号： G01N33/53 ,  G01N30/02

    CPC分类号： C12Q1/485 ,  C12N9/1205 ,  G01N33/57426 ,  G01N2333/91215 ,  G01N2500/02 ,  G01N2500/10

    摘要： The invention provides in a first aspect a method for the identification of an ZAP-70 interacting compound, comprising the steps of a) providing a protein preparation containing phosphorylated ZAP-70, b) contacting the protein preparation with aminopyrido-pyrimidine ligand 24 immobilized on a solid support under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex, c) incubating the aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex with a given compound, and d) determining whether the compound is able to separate phosphorylated ZAP-70 from the immobilized aminopyrido-pyrimidine ligand 24. In a second aspect, the present invention relates to A method for the identification of an ZAP-70 interacting compound, comprising the steps of a) providing a protein preparation containing phosphorylated ZAP-70, b) contacting the protein preparation with ligand aminopyrido-pyrimidine ligand 24 immobilized on a solid support and with a given compound under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex, and c) detecting the aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex formed in step b). In a third aspect, the invention provides a method for the identification of an ZAP-70 interacting compound, comprising the steps of: a) providing two aliquots of a protein preparation containing phosphorylated ZAP-70, b) contacting one aliquot with aminopyrido-pyrimidine ligand 24 immobilized on a solid support under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex, c) contacting the other aliquot with aminopyrido-pyrimidine ligand 24 immobilized on a solid support and with a given compound under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex, and d) determining the amount of aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex formed in steps b) and c). In a forth aspect, the invention relates to a method for the identification of an ZAP-70 interacting compound, comprising the steps of: a) providing two aliquots comprising each at least one cell containing phosphorylated ZAP-70, b) incubating one aliquot with a given compound, c) harvesting the cells of each aliquot, d) lysing the cells in order to obtain protein preparations, e) contacting the protein preparations with aminopyrido-pyrimidine ligand 24 immobilized on a solid support under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex, and f) determining the amount of aminopyrido-pyrimidine ligand 24—phosphorylated ZAP-70 complex formed in each aliquot in step e).