Luminescence-based methods and probes for measuring cytochrome P450 activity
    62.
    发明授权
    Luminescence-based methods and probes for measuring cytochrome P450 activity 有权
    用于测量细胞色素P450活性的基于发光的方法和探针

    公开(公告)号:US07692022B2

    公开(公告)日:2010-04-06

    申请号:US10665314

    申请日:2003-09-19

    IPC分类号: C07D277/68 C07D417/04

    摘要: The present invention provides methods, compositions, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferin or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. Upon addition of the luciferin derivative or other luminogenic molecule into a P450 reaction, the P450 enzyme metabolizes the molecule into a bioluminescent enzyme substrate, e.g., luciferin and/or luciferin derivative metabolite, in a P450 reaction. The resulting metabolite(s) serves as a substrate of the bioluminescent enzyme, e.g., luciferase, in a second light-generating reaction. Luminescent cytochrome P450 assays with low background signals and high sensitivity are disclosed and isoform selectivity is demonstrated. The present invention also provides an improved method for performing luciferase reactions which employs added pyrophosphatase to remove inorganic pyrophosphate, a luciferase inhibitor which may be present in the reaction mixture as a contaminant or may be generated during the reaction. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

    摘要翻译: 本发明提供了可用于分析细胞,组织和动物中的代谢活性以及筛选测试化合物对细胞色素P450活性的影响的方法,组合物,底物和试剂盒。 特别地,使用发光分子的一步和两步法。 荧光素或coelenterazines,其是细胞色素P450底物并且也是生物发光酶,例如萤光素酶,原底物。 在P450反应中加入荧光素衍生物或其他发光分子时,P450酶在P450反应中将分子代谢为生物发光酶底物,例如荧光素和/或荧光素衍生物代谢产物。 所产生的代谢物在第二次发光反应中用作生物发光酶,例如荧光素酶的底物。 公开了具有低背景信号和高灵敏度的发光细胞色素P450测定,证明了同工型选择性。 本发明还提供一种用于进行荧光素酶反应的改进方法,其使用添加的焦磷酸酶来除去无机焦磷酸盐,荧光素酶抑制剂,其可作为污染物存在于反应混合物中,或可能在反应期间产生。 本发明方法还提供了一种使用荧光素酶稳定剂如可逆荧光素酶抑制剂在基于荧光素酶的测定中稳定和延长发光信号的方法。

    ADP detection based luminescent phosphotransferase or ATP hydrolase assay
    63.
    发明申请
    ADP detection based luminescent phosphotransferase or ATP hydrolase assay 有权
    基于ADP检测的发光磷酸转移酶或ATP水解酶测定

    公开(公告)号:US20100075350A1

    公开(公告)日:2010-03-25

    申请号:US12460573

    申请日:2009-07-20

    IPC分类号: C12Q1/48 C12Q1/34 C12Q1/66

    CPC分类号: C12Q1/008

    摘要: The invention provides compositions and methods to determine or detect the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP.

    摘要翻译: 本发明提供了确定或检测酶的活性的组合物和方法,包括磷酸转移酶如激酶(例如蛋白质,脂质和糖激酶)和ATP水解酶例如使用ATP作为底物的ATP酶(例如HSP90)和形式 ADP作为产品,通过监测ADP的变化。

    Method for Detecting Transferase Enzymatic Activity
    65.
    发明申请
    Method for Detecting Transferase Enzymatic Activity 审中-公开
    检测转移酶酶活性的方法

    公开(公告)号:US20100021949A1

    公开(公告)日:2010-01-28

    申请号:US12561590

    申请日:2009-09-17

    IPC分类号: C12Q1/48

    CPC分类号: C12Q1/66 C12Q1/485

    摘要: Methods and kits for detecting transferase activity in a sample by measuring ATP using a composition comprising an ATP-dependent bioluminescence-generating enzyme, a luminogenic molecule and one or more transferase quenching agents.

    摘要翻译: 通过使用包含ATP依赖性生物发光酶,发光分子和一种或多种转移酶猝灭剂的组合物测量ATP来检测样品中转移酶活性的方法和试剂盒。

    Radio frequency identification method and system of distributing products
    66.
    发明授权
    Radio frequency identification method and system of distributing products 有权
    射频识别方法及系统分发产品

    公开(公告)号:US07591421B2

    公开(公告)日:2009-09-22

    申请号:US11247253

    申请日:2005-10-12

    IPC分类号: G06F7/08

    摘要: A method and system for vending products from a defined area, such as a micro-warehouse with a door. The method includes fitting each product with a radio frequency identification tag, positioning the plurality of products in a micro-warehouse, sensing opening and closing of the micro-warehouse door, scanning the plurality of products in the micro-warehouse upon sensing closing of the door to determine the number and type of products in the micro-warehouse, generating a message based on the number and type of products in the micro-warehouse, transmitting the message to a remote processor or server, and maintaining an inventory in the remote processor based on the message. The system is designed to be accessed by authorized individuals possessing some type of code or identifying mechanism. The micro-warehouses may be cabinets, refrigerators, secured rooms, or similar storage units or areas.

    摘要翻译: 用于从定义区域出售产品的方法和系统,例如具有门的微型仓库。 该方法包括使每个产品配有射频识别标签,将多个产品定位在微型仓库中,感测微型仓库门的打开和关闭,在感测关闭微型仓库门后扫描微型仓库中的多个产品 确定微型仓库中产品的数量和类型的门,根据微型仓库中的产品数量和类型生成消息,将消息传输到远程处理器或服务器,并在远程处理器中维护库存 基于消息。 该系统设计为由具有某种类型的代码或识别机制的授权人员访问。 微型仓库可以是机柜,冰箱,安全室或类似的存储单元或区域。

    SYNTHETIC NUCLEIC ACIDS FROM AQUATIC SPECIES
    67.
    发明申请
    SYNTHETIC NUCLEIC ACIDS FROM AQUATIC SPECIES 审中-公开
    来自水生物种的合成核酸

    公开(公告)号:US20090191622A1

    公开(公告)日:2009-07-30

    申请号:US12323270

    申请日:2008-11-25

    IPC分类号: C12N15/00 C12N15/11 C07H1/00

    摘要: A synthetic nucleic acid molecule is provided that includes nucleotides of a coding region for a fluorescent polypeptide having a codon composition differing at more than 25% of the codons from a parent nucleic acid sequence encoding a fluorescent polypeptide. The synthetic nucleic acid molecule has at least 3-fold fewer transcription regulatory sequences relative to the average number of such sequences in the parent nucleic acid sequence. The polypeptide encoded by the synthetic nucleic acid molecule preferably has at least 85% sequence identity to the polypeptide encoded by the parent nucleic acid sequence.

    摘要翻译: 提供了合成核酸分子,其包括编码区域的核苷酸,所述核苷酸的密码子组成不同于编码荧光多肽的母体核酸序列的密码子的25%以上的荧光多肽。 合成的核酸分子相对于母核酸序列中这样的序列的平均数目具有至少3倍的转录调节序列。 由合成核酸分子编码的多肽优选与由母体核酸序列编码的多肽具有至少85%的序列同一性。

    Methods of measuring luminogenic activity with a protected coelenterazine
    68.
    发明授权
    Methods of measuring luminogenic activity with a protected coelenterazine 有权
    用保护的肠培养液测量发光活性的方法

    公开(公告)号:US07537912B2

    公开(公告)日:2009-05-26

    申请号:US11832169

    申请日:2007-08-01

    IPC分类号: C12Q1/44

    摘要: A method of measuring the enzymatic activity of a luciferase includes contacting a luminogenic protein, such as a luciferase, with a protected luminophore to form a composition; and detecting light produced from the composition. The protected luminophore provides increased stability and improved signal-to-background ratios relative to the corresponding unmodified coelenterazine.

    摘要翻译: 测量荧光素酶的酶活性的方法包括将发光蛋白如荧光素酶与受保护的发光体接触以形成组合物; 并检测由该组合物产生的光。 受保护的发光体相对于相应的未修饰的肠培养基提供增加的稳定性和改善的信号与背景比。

    Treatment of sepsis
    69.
    发明授权
    Treatment of sepsis 失效
    治疗败血症

    公开(公告)号:US07494652B1

    公开(公告)日:2009-02-24

    申请号:US09095536

    申请日:1998-06-10

    申请人: John A. Kink

    发明人: John A. Kink

    IPC分类号: A61K39/395 C07K16/00

    摘要: Compositions and methods are described for treatment of sepsis in animals, including humans. Unique and specific combinations of polyclonal antibodies directed to cytokines are shown to have a beneficial effect in animal models predictive of human therapy.

    摘要翻译: 描述了用于治疗包括人在内的动物的败血症的组合物和方法。 显示针对细胞因子的多克隆抗体的独特和特异性组合在预测人类治疗的动物模型中具有有益效果。