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1. 发明公开

US20230416699A1 LUCIFERASE ENZYMES FOR USE WITH THERMOSTABLE LUCIFERINS IN BIOLUMINESCENT ASSAYS 审中-公开
公开(公告)号：US20230416699A1
公开(公告)日：2023-12-28
申请号：US18336636
申请日：2023-06-16
申请人： Promega Corporation
发明人： Michael Killoran , Ce Shi , Mary Hall , Lance P. Encell , Thomas Kirkland
IPC分类号： C12N9/02 , G01N21/76
CPC分类号： C12N9/0069 , C12Y113/12 , G01N21/763
摘要： Provided herein are enhanced luciferase enzymes for use with thermostable luciferin analogs for bioluminescent assays. In particular, the present disclosure provides compositions, assays, and methods for performing a bioluminescent assay using enhanced, high-activity luciferase enzymes compatible with thermostable luciferins, such as 5,5-disubstituted luciferin analogs.

2. 发明授权

US11365436B2 Synthetic oplophorus luciferases with enhanced light output 有权
公开(公告)号：US11365436B2
公开(公告)日：2022-06-21
申请号：US17075415
申请日：2020-10-20
申请人： Promega Corporation
发明人： Lance P. Encell , Mary Hall , Paul Otto , Gediminas Vidugiris , Keith V. Wood , Monika G. Wood , Kristopher Zimmerman
IPC分类号： C12Q1/66 , C12N9/00 , C12N9/02
摘要： A polynucleotide encoding a modified luciferase polypeptide. The modified luciferase polypeptide has at least 60% amino acid sequence identity to a wild-type Oplophorus luciferase and includes at least one amino acid substitution at a position corresponding to an amino acid in a wild-type Oplophorus luciferase of SEQ ID NO:1. The modified luciferase polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the wild-type Oplophorus luciferase.

3. 发明申请

US20190338340A1 5,5-DISUBSTITUTED LUCIFERINS AND THEIR USE IN LUCIFERASE-BASED ASSAYS 审中-公开
公开(公告)号：US20190338340A1
公开(公告)日：2019-11-07
申请号：US16516974
申请日：2019-07-19
申请人： Promega Corporation
发明人： Lance P. Encell , Mary P. Hall , Michael Killoran , Thomas A. Kirkland , Poncho Meisenheimer , Ce Shi
IPC分类号： C12Q1/66 , C07D513/04 , C07D513/10 , C07D277/66 , G01N21/76 , C07D417/04
摘要： Described are luciferin analogues, methods for making the analogues, kits including the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays.

4. 发明申请

US20190040449A1 COMPOSITIONS AND METHODS FOR STABILIZING BENZOTHIAZOLE LUCIFERIN ANALOGS 审中-公开
公开(公告)号：US20190040449A1
公开(公告)日：2019-02-07
申请号：US16053413
申请日：2018-08-02
申请人： PROMEGA CORPORATION
发明人： Ce Shi , Mary P. Hall , Thomas A. Kirkland , Lance P. Encell , Poncho Meisenheimer , Wenhui Zhou
IPC分类号： C12Q1/66 , C07D417/04
摘要： Disclosed herein are compositions and methods for stabilizing a benzothiazole luciferin analog such as D-luciferin and 6-amino-D-luciferin. The compositions may include the benzothiazole luciferin analog, a thionucleobase compound of formula (I), and a liquid medium, in which the thionucleobase is present in an amount effective to stabilize the luminogenic composition against decomposition. The methods provided herein may stabilize the benzothiazole luciferin analog against decomposition by contacting the benzothiazole luciferin analog with an effective amount of the thionucleobase compound in the presence of a liquid medium. Also provided herein is a kit containing the composition.

5. 发明申请

US20180327810A1 OPLOPHORUS-DERIVED LUCIFERASES, NOVEL COELENTERAZINE SUBSTRATES, AND METHODS OF USE 审中-公开
公开(公告)号：US20180327810A1
公开(公告)日：2018-11-15
申请号：US15948070
申请日：2018-04-09
申请人： Promega Corporation
发明人： Brock Binkowski , Lance P. Encell , Mary Hall , Matthew B. Robers , Michael R. Slater , Keith V. Wood , Monika G. Wood
IPC分类号： C12Q1/66 , C12N15/52 , C12N9/02 , C07D487/04
摘要： An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.

6. 发明申请

US20180155762A1 5,5-DISUBSTITUTED LUCIFERINS AND THEIR USE IN LUCIFERASE-BASED ASSAYS 审中-公开
公开(公告)号：US20180155762A1
公开(公告)日：2018-06-07
申请号：US15829581
申请日：2017-12-01
申请人： PROMEGA CORPORATION
发明人： Lance P. Encell , Mary P. Hall , Michael Killoran , Thomas A. Kirkland , Poncho Meisenheimer , Ce Shi
IPC分类号： C12Q1/66 , C07D417/04 , C07D513/04 , C07D513/10 , G01N21/76
CPC分类号： C12Q1/66 , C07D277/66 , C07D417/04 , C07D513/04 , C07D513/10 , G01N21/763
摘要： Described are luciferin analogues, methods for making the analogues, kits including the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays.

7. 发明授权

US09951373B2 Oplophorus-derived luciferases, novel coelenterazine substrates, and methods of use 有权
公开(公告)号：US09951373B2
公开(公告)日：2018-04-24
申请号：US14160282
申请日：2014-01-21
申请人： Promega Corporation
发明人： Brock Binkowski , Lance P. Encell , Mary Hall , Matthew B. Robers , Michael R. Slater , Keith V. Wood , Monika G. Wood
IPC分类号： C12N9/02 , C12N1/21 , C12N15/00 , C12Q1/66 , C07D487/04 , C12N15/52
CPC分类号： C12Q1/66 , A01K2217/072 , C07D487/04 , C12N9/0004 , C12N9/0069 , C12N15/52 , C12N2009/02 , C12Y113/12005 , C12Y113/12007
摘要： An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.

8. 发明授权

US09593316B2 Polynucleotides encoding mutant hydrolase proteins with enhanced kinetics and functional expression 有权
标题翻译：编码突变体水解酶蛋白的多核苷酸具有增强的动力学和功能表达
公开(公告)号：US09593316B2
公开(公告)日：2017-03-14
申请号：US14285327
申请日：2014-05-22
申请人： Promega Corporation
发明人： Aldis Darzins , Lance P. Encell , Rachel Friedman Ohana , Paul Otto , Michael R. Slater , Gediminas Vidugiris , Keith V. Wood , Monika G. Wood , Kate Qin Zhao , Kris Zimmerman
IPC分类号： C07H21/00 , C12N1/21 , C12N1/15 , C12N9/14 , C12Q1/34 , G01N33/58
CPC分类号： C12N9/14 , C12N9/96 , C12N11/00 , C12Q1/34 , C12Y308/01 , G01N33/581
摘要： The invention provides a mutant hydrolase protein with enhanced kinetics and functional expression, as well as polynucleotides encoding the mutant proteins and methods of using the polynucleotides and mutant proteins.
摘要翻译：本发明提供具有增强的动力学和功能表达的突变水解酶蛋白，以及编码突变蛋白的多核苷酸以及使用多核苷酸和突变蛋白的方法。

9. 发明授权

US09339561B2 Mutant protease biosensors with enhanced detection characteristics 有权
标题翻译：具有增强检测特性的突变蛋白酶生物传感器
公开(公告)号：US09339561B2
公开(公告)日：2016-05-17
申请号：US14269669
申请日：2014-05-05
申请人： Promega Corporation
发明人： Brock Binkowski , Braeden Butler , Lance P. Encell , Frank Fan , Brad Hook , Paul Otto , Gediminas Vidugiris , Susan Wigdal , Kristopher Zimmerman
IPC分类号： A61K49/00 , C12N9/02 , C12Q1/37 , C12Q1/66 , A01K67/027
CPC分类号： A61K49/0045 , A01K67/0275 , A61K49/0013 , C12N9/0069 , C12Q1/37 , C12Q1/66 , C12Y113/12007
摘要： A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.
摘要翻译：编码包含修饰的环状置换的热稳定性荧光素酶的生物传感器多肽的多核苷酸和将热稳定性荧光素酶的C末端部分连接到热稳定性萤光素酶的N末端部分的接头。相对于亲本循环置换的热稳定性荧光素酶，修饰的环状置换的热稳定性荧光素酶被修饰。接头包含能够与细胞中的靶分子相互作用的传感器区域。经修饰的环状置换的热稳定荧光素酶在目标分子存在下相对于亲本循环置换的热稳定性荧光素酶在生物传感器与靶分子相互作用后具有增强的响应。或者，修饰的环状置换的热稳定荧光素酶在生物传感器与目标分子相互作用后相对于在不存在目标分子的情况下相对于经修饰的环状置换的热稳定性荧光素酶具有增强的响应。

10. 发明授权

US09290794B2 Mutant protease biosensors with enhanced detection characteristics 有权
公开(公告)号：US09290794B2
公开(公告)日：2016-03-22
申请号：US13674655
申请日：2012-11-12
申请人： PROMEGA CORPORATION
发明人： Brock Binkowski , Braeden Butler , Lance P. Encell , Frank Fan , Brad Hook , Paul Otto , Kimberly Risma , Gediminas Vidugiris , Susan Wigdal , Kristopher Zimmerman
IPC分类号： C12Q1/66 , C12Q1/37 , C12N9/02
CPC分类号： C12Q1/66 , C12N9/0069 , C12Q1/37
摘要： A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

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