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21. 发明授权

US07157223B2 Method and apparatus for detecting enzymatic activity using molecules that change electrophoretic mobility 失效
标题翻译：使用改变电泳迁移率的分子检测酶活性的方法和装置
公开(公告)号：US07157223B2
公开(公告)日：2007-01-02
申请号：US09945396
申请日：2001-08-30
申请人： Nancy Allbritton , Christopher Sims , Michael W. Berns , Gavin D. Meredith , Tatiana B. Krasieva , Bruce J. Tromberg , Chao L. Lee
发明人： Nancy Allbritton , Christopher Sims , Michael W. Berns , Gavin D. Meredith , Tatiana B. Krasieva , Bruce J. Tromberg , Chao L. Lee
IPC分类号： C12Q1/00
CPC分类号： C12M41/46 , C12M35/02 , C12M47/06 , C12Q1/00 , C12Q1/485 , G01N27/44721 , G01N27/44743 , G01N33/50 , G01N33/5017
摘要： The activity of intracellular chemical reactions of molecules is measured by the use of fluorescently labeled substrate molecules that undergo a change in electrophoretic mobility upon chemical reaction such as that catalyzed by an enzyme. Specificity is achieved by using labeled substrate molecules that can be acted upon only by specific enzymes. Thus the activity of a specific enzyme or class of enzymes can be determined. Measurements are made with the intracellular presence of such substrate molecules, at some time of interest, typically after exposure of the cell to a stimulus that activates a particular enzymatic pathway. To ensure accuracy, measurements must be made in a timely manner so as to minimize chemical reactions occurring subsequent to the time of interest. Fast controllable laser lysis is used to obtain the contents of a single cell into which reporter substrate molecules have been introduced. The cell contents are then subjected to capillary electrophoresis and enzymatic activity is determined by comparing amounts of substrate molecules to amounts of enzymatically altered substrate molecules which are separated by the electrophoresis and identified by the presence of a fluorescent label.
摘要翻译：通过使用荧光标记的底物分子来测量分子的细胞内化学反应的活性，所述底物分子在诸如由酶催化的化学反应时经历电泳迁移率的变化。通过使用只能被特定酶作用的标记底物分子来实现特异性。因此，可以确定特异性酶或酶类的活性。通常在感兴趣的某个时间通常在将细胞暴露于激活特定酶促途径的刺激之后，通过细胞内存在这样的底物分子进行测量。为了确保准确性，必须及时进行测量，以最大限度地减少感兴趣的时间后发生的化学反应。使用快速可控的激光裂解来获得引入了报告基质分子的单细胞的含量。然后对细胞内容物进行毛细管电泳，并通过将底物分子的量与通过电泳分离并通过荧光标记的存在鉴定的酶促改变的底物分子的量进行比较来确定酶活性。

22. 发明申请

US20050148031A1 Catalytic efficiency and/or specificity of non-native substrates of enzymes 审中-公开
标题翻译：酶的非天然底物的催化效率和/或特异性
公开(公告)号：US20050148031A1
公开(公告)日：2005-07-07
申请号：US10754438
申请日：2004-01-09
申请人： Nancy Allbritton , Lee Bardwell , Christopher Sims
发明人： Nancy Allbritton , Lee Bardwell , Christopher Sims
IPC分类号： C07K1/13 , C07K14/47 , C07K16/40 , C12Q1/42 , C12Q1/48 , G01N33/50 , G01N33/53 , G01N33/567
CPC分类号： C07K1/13 , C07K2319/10 , C07K2319/23 , C07K2319/60 , C12Q1/42 , C12Q1/485 , G01N33/5005 , G01N2800/52
摘要： A method of introducing a fluorescent label into a cell, comprising: exposing a reporter to the cell, wherein the reporter comprises: a peptide substrate for an enzyme, a docking domain for the enzyme, attached to the peptide substrate, a membrane traversing moiety, and the label.
摘要翻译：一种将荧光标记引入细胞的方法，包括：将报告物暴露于细胞，其中所述报告基团包括：用于酶的肽底物，连接到肽底物的酶的对接结构域，穿膜部分的膜，和标签。

23. 发明申请

US20050282239A1 Cell-permeable enzyme activation reporter that can be loaded in a high throughput and gentle manner 审中-公开
标题翻译：细胞可渗透的酶激活报告物，可以高通量和温和的方式加载
公开(公告)号：US20050282239A1
公开(公告)日：2005-12-22
申请号：US11016542
申请日：2004-12-17
申请人： Nancy Allbritton , Christopher Sims , Francis Rossi , Joseph Soughayer
发明人： Nancy Allbritton , Christopher Sims , Francis Rossi , Joseph Soughayer
IPC分类号： C07K14/47 , C12Q1/48
CPC分类号： C12Q1/485 , C07K14/47
摘要： A membrane traversing peptide conjugate comprises (i) a label, (ii) a target peptide, attached to the label, and (iii) a transduction domain, attached to the target peptide. The target peptide is a reactant for a chemical reaction occurring in a cell.
摘要翻译：膜穿越肽缀合物包括（i）附着于标记的标记，（ii）靶肽，和（iii）与靶肽连接的转导结构域。靶肽是在细胞中发生化学反应的反应物。

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