公开(公告)号：US07238842B2

公开(公告)日：2007-07-03

申请号：US10768976

申请日：2004-01-30

申请人： Keith V. Wood ,  Dieter Klaubert ,  Georgyi V. Los ,  Robert F. Bulleit ,  Mark McDougall ,  Chad Zimprich

发明人： Keith V. Wood ,  Dieter Klaubert ,  Georgyi V. Los ,  Robert F. Bulleit ,  Mark McDougall ,  Chad Zimprich

IPC分类号： C07C22/00 ,  C08L67/00 ,  C07D235/02 ,  C12Q1/34

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。

公开(公告)号：US20070087400A1

公开(公告)日：2007-04-19

申请号：US11509796

申请日：2006-08-24

申请人： Aldis Darzins ,  Lance Encell ,  Tonny Johnson ,  Dieter Klaubert ,  Georgyi Los ,  Mark McDougall ,  Keith Wood ,  Monika Wood ,  Chad Zimprich

发明人： Aldis Darzins ,  Lance Encell ,  Tonny Johnson ,  Dieter Klaubert ,  Georgyi Los ,  Mark McDougall ,  Keith Wood ,  Monika Wood ,  Chad Zimprich

IPC分类号： C12Q1/26 ,  C12P21/06 ,  C07H21/04 ,  C12N9/16 ,  C12N9/88 ,  C12N1/21

CPC分类号： C12Q1/34 ,  B82Y30/00

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。

公开(公告)号：US09816054B2

公开(公告)日：2017-11-14

申请号：US12249584

申请日：2008-10-10

申请人： Sergei Saveliev ,  Daniel Simpson ,  Keith V. Wood ,  Harry Tetsuo Uyeda ,  Carolyn Woodroofe Hitko ,  Dieter Klaubert

发明人： Sergei Saveliev ,  Daniel Simpson ,  Keith V. Wood ,  Harry Tetsuo Uyeda ,  Carolyn Woodroofe Hitko ,  Dieter Klaubert

IPC分类号： C07C309/64 ,  C11D1/20 ,  C07C309/10 ,  C07C309/14 ,  C07C309/15 ,  C07C309/16 ,  C07C323/12 ,  C07C329/06 ,  C07D307/40 ,  C07D307/56 ,  C07D317/22 ,  C07D333/16 ,  C11D1/14

CPC分类号： C11D1/20 ,  C07C309/10 ,  C07C309/14 ,  C07C309/15 ,  C07C309/16 ,  C07C323/12 ,  C07C329/06 ,  C07D307/40 ,  C07D307/56 ,  C07D317/22 ,  C07D333/16 ,  C11D1/143

摘要： The invention provides surfactant compounds of formulas I-IX, which can be used in methods for aiding the solubilization, digestion, preparation, analysis, and/or characterization of biological material, for example, proteins or cell membranes. The compounds can also aid in the recovery of peptides generated during protein digestion, particularly for in-gel digestion protocol. Additionally, the compounds can improve enzymatic protein deglycosylation without interfering with downstream sample preparation steps and mass spectrometric analysis. The compounds can be specifically useful as digestion aids that can be decomposed by an acid, by heat, or a combination thereof. Decomposition of the surfactants allows for facile separation from isolated samples, and/or allows for analysis of the sample without interfering with the sensitivity of various analytical techniques.

公开(公告)号：US08895787B2

公开(公告)日：2014-11-25

申请号：US13450233

申请日：2012-04-18

申请人： Keith V. Wood ,  Georgyi V. Los ,  Robert F. Bulleit ,  Dieter Klaubert ,  Mark McDougall ,  Chad Zimprich

发明人： Keith V. Wood ,  Georgyi V. Los ,  Robert F. Bulleit ,  Dieter Klaubert ,  Mark McDougall ,  Chad Zimprich

IPC分类号： C07C19/00 ,  C07C21/00 ,  G01N33/53 ,  C12Q1/34 ,  C12N9/24

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。

公开(公告)号：US08765969B2

公开(公告)日：2014-07-01

申请号：US12217374

申请日：2008-07-03

申请人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel K. Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

发明人： James J. Cali ,  Dieter Klaubert ,  William Daily ,  Samuel K. Ho ,  Susan Frackman ,  Erika Hawkins ,  Keith V. Wood

IPC分类号： C07D417/04

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 invention provides compounds, compositions, methods, 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. luciferins or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. 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的化合物，组合物，方法，底物和试剂盒，并用于筛选测试化合物对细胞色素P450活性的影响。 特别地，使用发光分子的一步和两步法。 荧光素或coelenterazine，即细胞色素P450底物，并且也是生物发光酶，例如荧光素酶，原底物。 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。

公开(公告)号：US08466269B2

公开(公告)日：2013-06-18

申请号：US13435970

申请日：2012-03-30

申请人： Aldis Darzins ,  Lance Encell ,  Dieter Klaubert ,  Georgyi V. Los ,  Mark McDougall ,  Keith V. Wood ,  Monika G. Wood ,  Chad Zimprich

发明人： Aldis Darzins ,  Lance Encell ,  Dieter Klaubert ,  Georgyi V. Los ,  Mark McDougall ,  Keith V. Wood ,  Monika G. Wood ,  Chad Zimprich

IPC分类号： C07H21/04 ,  C12N9/14 ,  C12N1/00 ,  C12P21/02

CPC分类号： G01N33/581 ,  C07K7/06 ,  C07K16/00 ,  C07K2317/622 ,  C07K2319/43 ,  C07K2319/61 ,  C09B1/00 ,  C09B11/24 ,  C09B23/04 ,  C09B23/06 ,  C09B23/105 ,  C12N9/14 ,  C12Q1/34 ,  C12Y308/01005 ,  G01N33/573 ,  G01N2333/914 ,  Y10S530/825

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。

公开(公告)号：US08168405B2

公开(公告)日：2012-05-01

申请号：US13046297

申请日：2011-03-11

申请人： Aldis Darzins ,  Lance Encell ,  Dieter Klaubert ,  Georgyi V. Los ,  Mark McDougall ,  Keith V. Wood ,  Monika G. Wood ,  Chad Zimprich

发明人： Aldis Darzins ,  Lance Encell ,  Dieter Klaubert ,  Georgyi V. Los ,  Mark McDougall ,  Keith V. Wood ,  Monika G. Wood ,  Chad Zimprich

IPC分类号： G01N33/53 ,  C12Q1/34 ,  C12N9/14 ,  C12P21/02 ,  C07C19/00

CPC分类号： G01N33/581 ,  C07K7/06 ,  C07K16/00 ,  C07K2317/622 ,  C07K2319/43 ,  C07K2319/61 ,  C09B1/00 ,  C09B11/24 ,  C09B23/04 ,  C09B23/06 ,  C09B23/105 ,  C12N9/14 ,  C12Q1/34 ,  C12Y308/01005 ,  G01N33/573 ,  G01N2333/914 ,  Y10S530/825

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。

公开(公告)号：USRE42931E1

公开(公告)日：2011-11-15

申请号：US12284010

申请日：2008-09-17

申请人： Keith V. Wood ,  Dieter Klaubert ,  Georgyi V. Los ,  Robert F. Bulleit ,  Mark McDougall ,  Chad Zimprich

发明人： Keith V. Wood ,  Dieter Klaubert ,  Georgyi V. Los ,  Robert F. Bulleit ,  Mark McDougall ,  Chad Zimprich

IPC分类号： C07C22/00 ,  C08L67/00 ,  C07D235/02 ,  C12Q1/34

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。

公开(公告)号：US20100249427A1

公开(公告)日：2010-09-30

申请号：US12706078

申请日：2010-02-16

申请人： Martha O'Brien ,  Keith V. Wood ,  Dieter Klaubert ,  William Daily

发明人： Martha O'Brien ,  Keith V. Wood ,  Dieter Klaubert ,  William Daily

IPC分类号： C07D277/64

CPC分类号： C07D277/66 ,  C12Q1/37 ,  C12Q1/66

摘要： A sensitive bioluminescent assay to detect proteases including caspases, trypsin and tryptase is provided.

摘要翻译： 提供了敏感的生物发光测定法来检测蛋白酶，包括胱天蛋白酶，胰蛋白酶和类胰蛋白酶。

公开(公告)号：US07666987B2

公开(公告)日：2010-02-23

申请号：US11347054

申请日：2006-02-03

申请人： Martha O'Brien ,  Keith V. Wood ,  Dieter Klaubert ,  William Daily

发明人： Martha O'Brien ,  Keith V. Wood ,  Dieter Klaubert ,  William Daily

IPC分类号： C07K1/00

CPC分类号： C07D277/66 ,  C12Q1/37 ,  C12Q1/66

摘要： A sensitive bioluminescent assay to detect proteases including caspases is provided which employs an aminoluciferin or a carboxy-terminal protected derivative thereof covalently linked via a peptide bond to a substrate for a caspase or an aminoluciferin or a carboxy-terminal protected derivative thereof covalently linked via a peptide bond to a peptide substrate comprising aspartate that is specifically cleaved by a protease specific for the substrate.

摘要翻译： 提供了用于检测包括胱天蛋白酶的蛋白酶的敏感生物发光测定法，其使用通过肽键共价连接的氨基荧光素或其羧基端保护的衍生物与底物进行共价连接的胱天蛋白酶或氨基荧光素或羧基端保护的衍生物 肽结合到包含由对底物特异的蛋白酶特异性切割的天冬氨酸的肽底物。