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    Fluorescent capture assay for kinase activity employing anti-phosphotyrosine antibodies as capture and detection agents
    13.
    发明授权
    Fluorescent capture assay for kinase activity employing anti-phosphotyrosine antibodies as capture and detection agents 失效
    使用抗磷酸酪氨酸抗体作为捕获和检测剂的激酶活性的荧光捕获测定

    公开(公告)号：US06406869B1

    公开(公告)日：2002-06-18

    申请号：US09425549

    申请日：1999-10-22

    申请人： J. Fraser Glickman ,  James Inglese ,  Bassam Damaj

    发明人： J. Fraser Glickman ,  James Inglese ,  Bassam Damaj

    IPC分类号： G01N33543

    CPC分类号： G01N33/573 ,  C12Q1/48 ,  G01N2333/91235

    摘要： A method for determining the level of tyrosine kinase activity in a biological sample is disclosed. The method employs an anti-phosphotyrosine antibody as both the capture agent and the detecting agent. The detecting antibody is labeled with a lanthanide ion, such as europium, as the signal generating entity. The method is particularly well suited to high throughput screening, for example, for compounds which modulate tyrosine kinase activity.

    摘要翻译： 公开了确定生物样品中酪氨酸激酶活性水平的方法。 该方法采用抗磷酸酪氨酸抗体作为捕获剂和检测剂。 检测抗体用镧系离子如铕标记，作为信号发生实体。 该方法特别适用于高通量筛选，例如调节酪氨酸激酶活性的化合物。

    Biological assay detection method
    15.
    发明申请
    Biological assay detection method 审中-公开
    生物检测方法

    公开(公告)号：US20050164311A1

    公开(公告)日：2005-07-28

    申请号：US10484859

    申请日：2002-08-02

    申请人： James Inglese ,  Marc Ferrer ,  Aaron Hamilton

    发明人： James Inglese ,  Marc Ferrer ,  Aaron Hamilton

    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

    CPC分类号： G01N33/573 ,  G01N33/542

    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.

    摘要翻译： 本发明是检测反应产物诱导剂如酶的信号的反应产物的方法。 该方法能够识别构成检测策略基础的表位，而不需要针对表位的特异性抗体。 在该方法中，使用直接或间接标记的模块结构域和生物素化形式的同源肽配体作为可测量相互作用的基础。 可以通过例如Ser或Thr残基的磷酸化或氨基酸序列延伸超过C末端Val的修饰来掩蔽肽配体。 因为掩蔽的残基对于标记的模块结构域的结合至关重要，所以至少一个残基的掩蔽防止结合。 在通过适当的酶处理掩蔽的残留物（例如用磷酸酶处理磷酸化残基或用蛋白酶处理延长的残基时），将肽转化为能够结合到 标记的模块化领域并形成可衡量的复杂。

    Incorporation of phosphorylation sites
    17.
    发明授权
    Incorporation of phosphorylation sites 失效
    掺入磷酸化位点

    公开(公告)号：US06335176B1

    公开(公告)日：2002-01-01

    申请号：US09174216

    申请日：1998-10-16

    申请人： James Inglese ,  Joseph Fraser Glickman

    发明人： James Inglese ,  Joseph Fraser Glickman

    IPC分类号： C07K1113

    CPC分类号： C07K7/06 ,  C07K1/1072 ,  C07K1/113

    摘要： A reagent is described for incorporating phosphorylation sites into compounds, particularly into proteins and peptides. The reagent has the structure A—B—C wherein A is a moiety that is specifically reactive with a reactive side chain in the compound, B is a linking moiety, and C is a peptide sequence that contains a kinase substrate.

    摘要翻译： 描述了将磷酸化位点掺入化合物，特别是蛋白质和肽中的试剂。 试剂具有其中A是与化合物中的反应性侧链特异性反应的部分的结构，B是连接部分，C是含有激酶底物的肽序列。

    High-throughput assay
    18.
    发明授权
    High-throughput assay 失效
    高通量测定

    公开(公告)号：US5876946A

    公开(公告)日：1999-03-02

    申请号：US868280

    申请日：1997-06-03

    申请人： Jonathan J. Burbaum ,  Thomas D.Y. Chung ,  Gregory L. Kirk ,  James Inglese ,  Daniel Chelsky

    发明人： Jonathan J. Burbaum ,  Thomas D.Y. Chung ,  Gregory L. Kirk ,  James Inglese ,  Daniel Chelsky

    IPC分类号： G01N33/50 ,  C12Q1/25 ,  C40B30/04 ,  G01N33/15 ,  G01N33/543 ,  G01N33/68 ,  G01N33/53

    CPC分类号： C40B30/04 ,  C12Q1/25 ,  G01N33/54313 ,  G01N33/6845 ,  G01N33/6869 ,  G01N2500/00

    摘要： A homogeneous high throughput assay is described which screens compounds for enzyme inhibition, or receptor or other target binding. Inhibition (or binding) by the library compounds causes a change in the amount of an optically detectable label that is bound to suspendable cells or solid supports. The amounts of label bound to individual cells or solid supports are microscopically determined, and compared with the amount of label that is not bound to individual cells or solid supports. The degree of inhibition or binding is determined using this data. Confocal microscopy, and subsequent data analysis, allow the assay to be carried out without any separation step, and provide for high throughput screening of very small assay volumes using very small amounts of test compound.

    摘要翻译： 描述了筛选用于酶抑制的化合物或受体或其它靶结合的均匀高通量测定。 由文库化合物的抑制（或结合）导致与可悬浮细胞或固体支持物结合的光学可检测标记物的量的变化。 显微镜测定与单个细胞或固体支持物结合的标记物的量，并与不与单个细胞或固体支持物结合的标记物的量进行比较。 使用该数据确定抑制或结合的程度。 共聚焦显微镜和随后的数据分析允许在没有任何分离步骤的情况下进行测定，并且使用非常少量的测试化合物提供非常小的测定体积的高通量筛选。