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1. 发明申请

WO2006093529A3 COVALENT TETHERING OF FUNCTIONAL GROUPS TO PROTEINS AND SUBSTRATES THEREFOR 审中-公开
标题翻译：功能组合对蛋白质和底物的共价分析
公开(公告)号：WO2006093529A3
公开(公告)日：2007-03-22
申请号：PCT/US2005027307
申请日：2005-07-29
申请人： PROMEGA CORP , DARZINS ALDIS , ENCELL LANCE P , JOHNSON TONNY , KLAUBERT DIETER , LOS GEORGYI V , MCDOUGALL MARK , WOOD KEITH V , WOOD MONIKA G , ZIMPRICH CHAD
发明人： DARZINS ALDIS , ENCELL LANCE P , JOHNSON TONNY , KLAUBERT DIETER , LOS GEORGYI V , MCDOUGALL MARK , WOOD KEITH V , WOOD MONIKA G , ZIMPRICH CHAD
IPC分类号： G01N33/58 , C07K14/47 , C12N9/14 , C12Q1/26 , C12Q1/34
CPC分类号： C12N9/14 , B82Y5/00 , B82Y10/00 , B82Y30/00 , C07K2299/00 , C12N9/0069 , C12Q1/26 , C12Q1/34 , C12Q1/66 , C12Y113/12005 , C12Y308/01005
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译：提供任选融合到感兴趣的蛋白质的突变体水解酶。突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定，并且相对于野生型水解酶具有至少两个氨基酸取代野生型水解酶。还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。

2. 发明申请

WO2004072232A9 COVALENT TETHERING OF FUNCTIONAL GROUPS TO PROTEINS 审中-公开
标题翻译：功能团与蛋白质的共价结合
公开(公告)号：WO2004072232A9
公开(公告)日：2004-10-14
申请号：PCT/US2004002607
申请日：2004-01-30
申请人： PROMEGA CORP , WOOD KEITH V , LOS GEORGYI V , BULLEIT ROBERT F , KLAUBERT DIETER , MCDOUGALL MARK , ZIMPRICH CHAD
发明人： WOOD KEITH V , LOS GEORGYI V , BULLEIT ROBERT F , KLAUBERT DIETER , MCDOUGALL MARK , ZIMPRICH CHAD
IPC分类号： C12N20060101 , C12N9/14 , C12N9/86 , C12Q1/26 , C12Q1/34 , G01N33/50 , G01N33/58 , C12N
CPC分类号： C12Q1/34 , C07C217/08 , C07C271/16 , C07D209/48 , C07D233/64 , C07D311/78 , C07D311/82 , C07D405/06 , C07D493/10 , C07D495/04 , C07F5/022 , C12N9/14 , C12N9/86 , C12Y308/01005 , G01N33/5005 , G01N33/58 , Y10S436/80
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译：提供了任选地与感兴趣的蛋白质融合的突变水解酶。突变水解酶能够与相应的非突变型（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定。还提供了包含一个或多个官能团的用于水解酶的底物，以及使用突变水解酶和本发明底物的方法。还提供了能够与底物和表达融合蛋白的细胞形成稳定键的融合蛋白。

3. 发明申请

WO2004072232A3 COVALENT TETHERING OF FUNCTIONAL GROUPS TO PROTEINS 审中-公开
标题翻译：功能组合蛋白质的共价分析
公开(公告)号：WO2004072232A3
公开(公告)日：2005-01-27
申请号：PCT/US2004002607
申请日：2004-01-30
申请人： PROMEGA CORP , WOOD KEITH V , LOS GEORGYI V , BULLEIT ROBERT F , KLAUBERT DIETER , MCDOUGALL MARK , ZIMPRICH CHAD
发明人： WOOD KEITH V , LOS GEORGYI V , BULLEIT ROBERT F , KLAUBERT DIETER , MCDOUGALL MARK , ZIMPRICH CHAD
IPC分类号： C12N20060101 , C12N9/14 , C12N9/86 , C12Q1/26 , C12Q1/34 , G01N33/50 , G01N33/58
CPC分类号： C12Q1/34 , C07C217/08 , C07C271/16 , C07D209/48 , C07D233/64 , C07D311/78 , C07D311/82 , C07D405/06 , C07D493/10 , C07D495/04 , C07F5/022 , C12N9/14 , C12N9/86 , C12Y308/01005 , G01N33/5005 , G01N33/58 , Y10S436/80
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译：提供任选融合到感兴趣的蛋白质的突变体水解酶。突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定。还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。

4. 发明申请

WO2008118445A9 METHODS TO QUENCH LIGHT FROM OPTICAL REACTIONS 审中-公开
标题翻译：从光学反应中排除光的方法
公开(公告)号：WO2008118445A9
公开(公告)日：2008-12-04
申请号：PCT/US2008003924
申请日：2008-03-26
申请人： PROMEGA CORP , DAILY WILLIAM , HAWKINS ERIKA , KLAUBERT DIETER , MCDOUGALL MARK , UNCH JAMES , WOOD KEITH V , ZHOU WENHUI , ZHU JI
发明人： DAILY WILLIAM , HAWKINS ERIKA , KLAUBERT DIETER , MCDOUGALL MARK , UNCH JAMES , WOOD KEITH V , ZHOU WENHUI , ZHU JI
IPC分类号： C12Q1/66 , C07C211/30
CPC分类号： C12Q1/66
摘要： The present invention relates to single and dual reporter luminescence assays utilizing reagents to quench an optical, e.g., an enzyme-mediated luminescence, reaction. In one embodiment of the invention, a reagent is added to an assay which selectively quenches a first enzyme-mediated luminescence reaction without affecting a subsequent distinct enzyme-mediated luminescent reaction(s). An assay kit containing one or more selective quench reagents, and compositions comprising the quench reagent(s), are also provided.
摘要翻译：本发明涉及使用试剂淬灭光学例如酶介导的发光反应的单和双重报道发光测定。在本发明的一个实施方案中，将试剂加入到选择性淬灭第一酶介导的发光反应而不影响随后不同的酶介导的发光反应的测定中。还提供了含有一种或多种选择性骤冷试剂和包含骤冷试剂的组合物的测定试剂盒。

5. 发明申请

WO2006130551A8 LUMINOGENIC AND FLUOROGENIC COMPOUNDS AND METHODS TO DETECT MOLECULES OR CONDITIONS 审中-公开
标题翻译：荧光和荧光化合物及检测分子或条件的方法
公开(公告)号：WO2006130551A8
公开(公告)日：2007-02-01
申请号：PCT/US2006020731
申请日：2006-05-31
申请人： PROMEGA CORP , DAILY WILLIAM , HAWKINS ERIKA , KLAUBERT DIETER , LIU JIANQUAN , MEISENHEIMER PONCHO , SCURRIA MICHAEL , SHULTZ JOHN W , UNCH JAMES , WOOD KEITH V , ZHOU WENHUI , VALLEY MICHAEL P , CALI JAMES J
发明人： DAILY WILLIAM , HAWKINS ERIKA , KLAUBERT DIETER , LIU JIANQUAN , MEISENHEIMER PONCHO , SCURRIA MICHAEL , SHULTZ JOHN W , UNCH JAMES , WOOD KEITH V , ZHOU WENHUI , VALLEY MICHAEL P , CALI JAMES J
IPC分类号： C12Q1/66 , C07D277/66
CPC分类号： C07D277/66 , C07D277/68 , C07D311/16 , C07D417/14 , C07D493/10 , C07F9/65583 , C09B11/24 , C09B19/00 , C09B57/00 , C09B57/02 , C12Q1/66 , G01N2500/00
摘要： A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided.
摘要翻译：提供了检测采用荧光素衍生物或荧光团衍生物的样品中至少一种分子的存在或量的方法。

6. 发明申请

WO2006130551A3 LUMINOGENIC AND FLUOROGENIC COMPOUNDS AND METHODS TO DETECT MOLECULES OR CONDITIONS 审中-公开
标题翻译：荧光和荧光化合物和检测分子或条件的方法
公开(公告)号：WO2006130551A3
公开(公告)日：2007-05-03
申请号：PCT/US2006020731
申请日：2006-05-31
申请人： PROMEGA CORP , DAILY WILLIAM , HAWKINS ERIKA , KLAUBERT DIETER , LIU JIANQUAN , MEISENHEIMER PONCHO , SCURRIA MICHAEL , SHULTZ JOHN W , UNCH JAMES , WOOD KEITH V , ZHOU WENHUI , VALLEY MICHAEL P , CALI JAMES J
发明人： DAILY WILLIAM , HAWKINS ERIKA , KLAUBERT DIETER , LIU JIANQUAN , MEISENHEIMER PONCHO , SCURRIA MICHAEL , SHULTZ JOHN W , UNCH JAMES , WOOD KEITH V , ZHOU WENHUI , VALLEY MICHAEL P , CALI JAMES J
IPC分类号： C12Q1/66 , C07D277/66
CPC分类号： C07D277/66 , C07D277/68 , C07D311/16 , C07D417/14 , C07D493/10 , C07F9/65583 , C09B11/24 , C09B19/00 , C09B57/00 , C09B57/02 , C12Q1/66 , G01N2500/00
摘要： A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided.
摘要翻译：提供了检测采用荧光素衍生物或荧光团衍生物的样品中至少一种分子的存在或量的方法。

7. 发明申请

WO2004027378A3 LUMINESCENCE-BASED METHODS AND PROBES FOR MEASURING CYTOCHROME P450 ACTIVITY 审中-公开
标题翻译：用于测量细胞色素P450活性的基于荧光的方法和探针
公开(公告)号：WO2004027378A3
公开(公告)日：2004-11-25
申请号：PCT/US0329078
申请日：2003-09-16
申请人： PROMEGA CORP , CALI JAMES J , KLAUBERT DIETER , DAILY WILLIAM , HO SAMUEL KIN SANG , FRACKMAN SUSAN , HAWKINS ERIKA , WOOD KEITH V
发明人： CALI JAMES J , KLAUBERT DIETER , DAILY WILLIAM , HO SAMUEL KIN SANG , FRACKMAN SUSAN , HAWKINS ERIKA , WOOD KEITH V
IPC分类号： C07D403/04 , C07D413/04 , C07D417/04 , C07D417/14 , C07F9/06 , C12N9/02 , C12Q1/00 , C12Q1/26 , C12Q1/66 , G01N20060101 , G01N33/53 , G01N33/58
CPC分类号： G01N33/502 , A61K49/0008 , C07D417/04 , C12Q1/66 , G01N33/5088 , G01N33/582 , G01N2333/90245 , G01N2333/90251 , G01N2500/10
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 lightgenerating 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。在第二次发光反应中，所得的代谢物用作生物发光酶，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。本发明的方法还提供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。

8. 发明申请

WO2011026085A3 REACTIVE CYANINE COMPOUNDS 审中-公开
标题翻译：反应性氰基化合物
公开(公告)号：WO2011026085A3
公开(公告)日：2011-04-21
申请号：PCT/US2010047274
申请日：2010-08-31
申请人： PROMEGA CORP , CORONA CESEAR , KLAUBERT DIETER , MCDOUGALL MARK
发明人： CORONA CESEAR , KLAUBERT DIETER , MCDOUGALL MARK
IPC分类号： C09B23/00 , C09B23/06 , C09B23/08 , C12Q1/68 , G01N21/64 , G01N33/58
CPC分类号： G01N33/582 , A61K49/0032 , A61K49/0056 , C09B23/0008 , C09B23/0066 , C09B23/06 , C09B23/083
摘要： The invention provides compounds and compositions of Formulas I-VII, and methods of using the compounds. The compounds can be used to prepare dye conjugates that are uniformly and substantially more fluorescent on proteins, nucleic acids or other biopolymers, than conjugates labeled with structurally similar known carbocyanine dyes. In addition to having more intense fluorescence emission than structurally similar dyes at virtually identical wavelengths, and decreased artifacts in their absorption spectra upon conjugation to biopolymers, the compounds can have greater photostability and/or higher absorbance (extinction coefficients) at the wavelength(s) of peak absorbance than such structurally similar dyes.
摘要翻译：本发明提供了式I-VII的化合物和组合物，以及使用该化合物的方法。该化合物可用于制备在蛋白质，核酸或其他生物聚合物上均匀且基本上更多荧光的染料缀合物，而不是与结构上相似的已知碳菁染料标记的缀合物。除了与实际相同的波长下的结构相似的染料具有更强烈的荧光发射以及在与生物聚合物共轭时其吸收光谱中的伪影减少，化合物在波长处具有更大的光稳定性和/或更高的吸光度（消光系数）的吸光度比这种结构相似的染料高。

9. 发明专利

JP2013208136A Mutant dehalogenase with increased binding stability 审中-公开
标题翻译：具有增加的结合稳定性的突变体
公开(公告)号：JP2013208136A
公开(公告)日：2013-10-10
申请号：JP2013131105
申请日：2013-06-21
申请人： Promega Corp , プロメガコーポレイションPromega Corporation
发明人： DARZINS ALDIS , ENCELL LANCE P , JOHNSON TONNY , KLAUBERT DIETER , LOS GEORGYI V , MCDOUGALL MARK , WOOD KEITH V , WOOD MONIKA G , ZIMBRICH CHAD
IPC分类号： C12N15/09 , C07K14/47 , C12N9/04 , C12N9/14 , C12Q1/26
CPC分类号： C12N9/14 , B82Y5/00 , B82Y10/00 , B82Y30/00 , C07K2299/00 , C12N9/0069 , C12Q1/26 , C12Q1/34 , C12Q1/66 , C12Y113/12005 , C12Y308/01005
摘要： PROBLEM TO BE SOLVED: To provide mutant proteins covalently linked (tethered) to one or more functional groups and methods for their use, and a mutant dehalogenase optionally fused to a protein of interest.SOLUTION: One or more functional groups are tethered (linked) to a protein to a fusion protein (chimera) which includes a protein via a covalent bond or otherwise stable bond. The mutant dehalogenase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) dehalogenase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has two amino acid substitutions relative to the wild-type dehalogenase.
摘要翻译：要解决的问题：提供共价连接（连接）到一个或多个官能团的突变蛋白及其使用方法，以及任选地与感兴趣的蛋白质融合的突变体脱卤酶。解决方案：将一个或多个官能团连接（连接）涉及包含通过共价键或其他稳定键的蛋白质的融合蛋白（嵌合体）的蛋白质。突变体脱卤酶能够与相应的非突变（野生型）脱卤素酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定，并且相对于野生型水解酶具有两个氨基酸取代，型脱卤素酶。

10. 发明专利

JP2008281575A Luminescence-based method and probe for measuring cytochrome p450 activity 有权
标题翻译：用于测量细胞色素P450活性的基于荧光的方法和探针
公开(公告)号：JP2008281575A
公开(公告)日：2008-11-20
申请号：JP2008146920
申请日：2008-06-04
申请人： Promega Corp , プロメガコーポレイションPromega Corporation
发明人： CALI JAMES J , KLAUBERT DIETER , DAILY WILLIAM , HO SAMUEL KIN SANG , FRACKMAN SUSAN , HAWKINS ERIKA , WOOD KEITH V
IPC分类号： G01N21/76 , C07D277/66 , C07D403/04 , C07D413/04 , C07D417/04 , C07D417/14 , C07F9/06 , C12N9/02 , C12Q1/00 , C12Q1/02 , C12Q1/26 , C12Q1/66 , C12Q1/68 , G01N20060101 , G01N33/53 , G01N33/58
CPC分类号： G01N33/502 , A61K49/0008 , C07D417/04 , C12Q1/66 , G01N33/5088 , G01N33/582 , G01N2333/90245 , G01N2333/90251 , G01N2500/10
摘要： PROBLEM TO BE SOLVED: To provide 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. SOLUTION: 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. COPYRIGHT: (C)2009,JPO&INPIT
摘要翻译：要解决的问题：提供可用于分析细胞，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活性影响的方法，组合物，底物和试剂盒。提供了使用作为细胞色素P450底物并且也是生物发光酶，例如荧光素酶，原底物的发光分子（例如荧光素或coelenterazine）的一步和两步方法。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光产生反应中用作生物发光酶，例如荧光素酶的底物。版权所有（C）2009，JPO＆INPIT

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