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1. 发明授权

US06893827B1 Receptor function assay for G-protein coupled receptors and orphan receptors by reporter enzyme mutant complementation 失效
标题翻译：通过报告酶突变体互补对G蛋白偶联受体和孤儿受体的受体功能测定
公开(公告)号：US06893827B1
公开(公告)日：2005-05-17
申请号：US09654499
申请日：2000-09-01
申请人： Michelle A. J. Palmer , Melissa Gee , Bonnie Tillotson , Xiao-Jia Chang
发明人： Michelle A. J. Palmer , Melissa Gee , Bonnie Tillotson , Xiao-Jia Chang
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12P21/04 , C12Q1/02 , C12Q1/34 , G01N33/566 , C07K14/705 , C07K19/00 , C12N15/62 , G01N33/567
CPC分类号： G01N33/566 , G01N2333/726 , G01N2500/10
摘要： Methods for detecting G-protein coupled receptor (GPCR) activity; methods of assaying GPCR activity; and methods of screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described.
摘要翻译：检测G蛋白偶联受体（GPCR）活性的方法; 测定GPCR活性的方法; 并描述了筛选GPCR配体，G蛋白偶联受体激酶（GRK）活性和与GPCR调节过程的组分相互作用的化合物的方法。

2. 发明授权

US07235374B2 Systems for sensitive detection of G-protein coupled receptor and orphan receptor function using reporter enzyme mutant complementation 有权
标题翻译：使用报告酶突变体互补对G蛋白偶联受体和孤儿受体功能敏感检测的系统
公开(公告)号：US07235374B2
公开(公告)日：2007-06-26
申请号：US10959611
申请日：2004-10-05
申请人： Michelle A. J. Palmer , Melissa Gee , Bonnie Tillotson , Xiao-jia Chang
发明人： Michelle A. J. Palmer , Melissa Gee , Bonnie Tillotson , Xiao-jia Chang
IPC分类号： C07K14/705 , C07K19/00 , C12N15/62 , G01N33/566
CPC分类号： G01N33/566 , G01N2333/726 , G01N2500/10
摘要： Methods for detecting G-protein coupled receptor (GPCR) activity; methods for assaying GPCR activity; and methods for screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described. Included are methods for expanding ICAST technologies for assaying GPCR activity with applications for ligand fishing, and agonist or antagonist screening. These methods include: engineering seronine/threonine phosphorylation sites into known or orphan GPCR open reading frames in order to increase the affinity of arrestin for the activated form of the GPCR or to increase the reside time of arrestin on the activated GPCR; engineering mutant arrestin proteins that bind to activated GPCRs in the absence of G-protein coupled receptor kinases which may be limiting; and engineering mutant super arrestin proteins that have an increased affinity for activated GPCRs with or without phosphorylation. These methods are intended to increase the robustness of the GPCR/ICAST technology in situations in which G-protein coupled receptor kinases are absent or limiting, or in which the GPCR is not efficiently down-regulated or is rapidly resensitized (thus having a labile interaction with arrestin). Included are also more specific methods for using ICAST complementary enzyme fragments to monitor GPCR homo- and hetero-dimerization with applications for drug lead discovery and ligand and function discovery for orphan GPCRs.
摘要翻译：检测G蛋白偶联受体（GPCR）活性的方法; 测定GPCR活性的方法; 并描述了筛选GPCR配体，G蛋白偶联受体激酶（GRK）活性和与GPCR调节过程的组分相互作用的化合物的方法。包括用于扩展用于测定GPCR活性的ICAST技术的方法，用于配体捕鱼和激动剂或拮抗剂筛选。这些方法包括：将已知或孤儿GPCR开放阅读框中的氨基酸/苏氨酸磷酸化位点工程化，以增加抑制蛋白对GPCR活化形式的亲和力或增加抑制蛋白在激活的GPCR上的停留时间; 在不存在可能是限制性的G蛋白偶联受体激酶的情况下结合激活的GPCR的工程化突变抑制蛋白; 和具有增加对具有或不具有磷酸化作用的活化GPCR的亲和力的工程突变超级抑制蛋白。这些方法旨在增加GPCR / ICAST技术在G蛋白偶联受体激酶不存在或限制的情况下的稳健性，或其中GPCR不能有效下调或快速复敏（因此具有不稳定相互作用与arrestin）。包括也是使用ICAST互补酶片段监测GPCR同源和异二聚化的更具体的方法，用于药物铅发现和孤儿GPCR的配体和功能发现的应用。

3. 发明授权

US06800445B2 Systems for sensitive detection of G-protein coupled receptor and orphan receptor function using reporter enzyme mutant complementation 失效
标题翻译：使用报告酶突变体互补对G蛋白偶联受体和孤儿受体功能敏感检测的系统
公开(公告)号：US06800445B2
公开(公告)日：2004-10-05
申请号：US09759152
申请日：2001-01-16
申请人： Michelle A. J. Palmer , Melissa Gee , Bonnie Tillotson , Xiao-jia Chang
发明人： Michelle A. J. Palmer , Melissa Gee , Bonnie Tillotson , Xiao-jia Chang
IPC分类号： G01N33567
CPC分类号： G01N33/566 , G01N2333/726 , G01N2500/10
摘要： Methods for detecting G-protein coupled receptor (GPCR) activity; methods for assaying GPCR activity; and methods for screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described. Included are methods for expanding ICAST technologies for assaying GPCR activity with applications for ligand fishing, and agonist or antagonist screening. These methods include: engineering seronine/threonine phosphorylation sites into known or orphan GPCR open reading frames in order to increase the affinity of arrestin for the activated form of the GPCR or to increase the reside time of arrestin on the activated GPCR; engineering mutant arrestin proteins that bind to activated GPCRs in the absence of G-protein coupled receptor kinases which may be limiting; and engineering mutant super arrestin proteins that have an increased affinity for activated GPCRs with or without phosphorylation. These methods are intended to increase the robustness of the GPCR/ICAST technology in situations in which G-protein coupled receptor kinases are absent or limiting, or in which the GPCR is not efficiently down-regulated or is rapidly resensitized (thus having a labile interaction with arrestin). Included are also more specific methods for using ICAST complementary enzyme fragments to monitor GPCR homo- and hetero-dimerization with applications for drug lead discovery and ligand and function discovery for orphan GPCRs.
摘要翻译：检测G蛋白偶联受体（GPCR）活性的方法; 测定GPCR活性的方法; 并描述了筛选GPCR配体，G蛋白偶联受体激酶（GRK）活性和与GPCR调节过程的组分相互作用的化合物的方法。包括用于扩展用于测定GPCR活性的ICAST技术的方法，用于配体捕鱼和激动剂或拮抗剂筛选。这些方法包括：将已知或孤儿GPCR开放阅读框中的氨基酸/苏氨酸磷酸化位点工程化，以增加抑制蛋白对GPCR活化形式的亲和力或增加抑制蛋白在激活的GPCR上的停留时间; 在不存在可能是限制性的G蛋白偶联受体激酶的情况下结合激活的GPCR的工程化突变抑制蛋白; 和具有增加对具有或不具有磷酸化作用的活化GPCR的亲和力的工程突变超级抑制蛋白。这些方法旨在增加GPCR / ICAST技术在G蛋白偶联受体激酶不存在或限制的情况下的稳健性，或其中GPCR不能有效下调或快速复敏（因此具有不稳定相互作用与arrestin）。包括也是使用ICAST互补酶片段监测GPCR同源和异二聚化的更具体的方法，用于药物铅发现和孤儿GPCR的配体和功能发现的应用。

4. 发明申请

US20060040250A1 Systems for sensitive detection of G-protein coupled receptor and orphan receptor function using reporter enzyme mutant complementation 有权
公开(公告)号：US20060040250A1
公开(公告)日：2006-02-23
申请号：US10959611
申请日：2004-10-05
申请人： Michelle Palmer , Melissa Gee , Bonnie Tillotson , Xiao-jia Chang
发明人： Michelle Palmer , Melissa Gee , Bonnie Tillotson , Xiao-jia Chang
IPC分类号： C12Q1/00 , G01N33/567 , G01N33/53
CPC分类号： G01N33/566 , G01N2333/726 , G01N2500/10
摘要： Methods for detecting G-protein coupled receptor (GPCR) activity; methods for assaying GPCR activity; and methods for screening for GPCR ligands, G-protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process are described. Included are methods for expanding ICAST technologies for assaying GPCR activity with applications for ligand fishing, and agonist or antagonist screening. These methods include: engineering seronine/threonine phosphorylation sites into known or orphan GPCR open reading frames in order to increase the affinity of arrestin for the activated form of the GPCR or to increase the reside time of arrestin on the activated GPCR; engineering mutant arrestin proteins that bind to activated GPCRs in the absence of G-protein coupled receptor kinases which may be limiting; and engineering mutant super arrestin proteins that have an increased affinity for activated GPCRs with or without phosphorylation. These methods are intended to increase the robustness of the GPCR/ICAST technology in situations in which G-protein coupled receptor kinases are absent or limiting, or in which the GPCR is not efficiently down-regulated or is rapidly resensitized (thus having a labile interaction with arrestin). Included are also more specific methods for using ICAST complementary enzyme fragments to monitor GPCR homo- and hetero-dimerization with applications for drug lead discovery and ligand and function discovery for orphan GPCRs.

5. 发明授权

US10031142B2 In situ chemiluminescent substrates and assays 有权
公开(公告)号：US10031142B2
公开(公告)日：2018-07-24
申请号：US13808922
申请日：2011-07-06
申请人： Alison Sparks , Zhixian Wang , Melissa Gee
发明人： Alison Sparks , Zhixian Wang , Melissa Gee
IPC分类号： G01N33/58 , C12Q1/42 , C12Q1/34 , C07F9/12 , C07F9/40 , C07F9/6541 , C07F9/655 , C07F9/6558 , C07H15/203 , C12Q1/6816 , C07D321/00
摘要： Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed.

6. 发明申请

US20120100566A1 CHEMILUMINESCENT COMPOSITIONS, METHODS, ASSAYS AND KITS FOR OXIDATIVE ENZYMES 有权
标题翻译：氧化酶的显色组合物，方法，测定和试剂
公开(公告)号：US20120100566A1
公开(公告)日：2012-04-26
申请号：US13254154
申请日：2010-03-01
申请人： Rouh-Rong Juo , Brooks Edwards , Melissa Gee , Zhixian Wang , Kathleen Skaare
发明人： Rouh-Rong Juo , Brooks Edwards , Melissa Gee , Zhixian Wang , Kathleen Skaare
IPC分类号： C12Q1/26 , C07D409/04 , C07D323/00
CPC分类号： C12Q1/26 , C07D305/14 , C07D321/00 , C07D323/00 , C07D407/04 , C07D409/04 , C07D493/04 , C07D493/10 , G01N2333/90245 , G01N2333/90251 , G01N2333/90258
摘要： Chemiluminescent compositions, methods, assays and kits for oxidative enzymes are described. Further disclosed are dioxetane compounds of the form: 0-0 ΛR R R T (i) where R can independently be any branched alkyl or cycloalkyl group which provides stabilization for the dioxetane or where both R groups together form a cycloalkyl or polycycloalkyl moiety spiro bound to the dioxetane ring, wherein each R group or the spiro bound moiety can be unsubstituted or substituted with one or more electron-withdrawing groups or electron donating groups, or groups providing preferential oxidative isozyme substrate recognition, and wherein Ri is an aryl group, or an alkyl group of 1-20 carbon atoms, which can be optionally substituted with 1 or more halogen atoms, and wherein T is an aryl or heteroaryl ring capable of emitting light upon enzyme activated decomposition of the dioxetane I. Kits, methods and assays are also disclosed that comprise the dioxetane compounds.
摘要翻译：描述了用于氧化酶的化学发光组合物，方法，测定和试剂盒。进一步公开的是以下形式的二氧杂环丁烷化合物：其中R可以独立地是任何分支烷基或环烷基，其为二氧杂环丁烷提供稳定性，或者其中两个R基团一起形成环烷基或多环烷基部分螺二环氧环乙烷环，其中每个R基团或螺环结合的部分可以是未取代的或被一个或多个吸电子基团或给电子基团或提供优先氧化同功酶底物识别的基团取代，并且其中R 1是芳基或烷基 1-20个碳原子的基团，其可以任选地被1个或多个卤素原子取代，并且其中T是能够在二氧杂环丁烷I的酶活化分解时发光的芳基或杂芳基环。另外还公开了方法和测定其包含二氧杂环丁烷化合物。

7. 发明授权

US09067910B2 Chemiluminescent compositions, methods, assays and kits for oxidative enzymes 有权
标题翻译：用于氧化酶的化学发光组合物，方法，测定和试剂盒
公开(公告)号：US09067910B2
公开(公告)日：2015-06-30
申请号：US13254154
申请日：2010-03-01
申请人： Rouh-Rong Juo , Brooks Edwards , Melissa Gee , Zhixian Wang , Kathleen Skaare
发明人： Rouh-Rong Juo , Brooks Edwards , Melissa Gee , Zhixian Wang , Kathleen Skaare
IPC分类号： C07D305/14 , C07D493/00 , C07D321/00 , C07D493/10 , C07D493/04 , C07D407/04 , C12Q1/26
CPC分类号： C12Q1/26 , C07D305/14 , C07D321/00 , C07D323/00 , C07D407/04 , C07D409/04 , C07D493/04 , C07D493/10 , G01N2333/90245 , G01N2333/90251 , G01N2333/90258
摘要： Chemiluminescent compositions, methods, assays and kits for oxidative enzymes are described. Further disclosed are dioxetane compounds of the form: where R can independently be any branched alkyl or cycloalkyl group which provides stabilization for the dioxetane or where both R groups together form a cycloalkyl or polycycloalkyl moiety spiro bound to the dioxetane ring, wherein each R group or the spiro bound moiety can be unsubstituted or substituted with one or more electron-withdrawing groups or electron donating groups, or groups providing preferential oxidative isozyme substrate recognition, and wherein R1 is an aryl group, or an alkyl group of 1-20 carbon atoms, which can be optionally substituted with 1 or more halogen atoms, and wherein T is an aryl or heteroaryl ring capable of emitting light upon enzyme activated decomposition of the dioxetane I. Kits, methods and assays are also disclosed that comprise the dioxetane compounds.
摘要翻译：描述了用于氧化酶的化学发光组合物，方法，测定和试剂盒。进一步公开的是以下形式的二氧杂环丁烷化合物：其中R可以独立地为任何为二氧杂环丁烷提供稳定化的支链烷基或环烷基，或者其中两个R基团一起形成与二氧杂环丁烷环结合的环烷基或多环烷基部分，其中每个R基团或螺结合部分可以是未取代的或被一个或多个吸电子基团或给电子基团或提供优先氧化同功酶底物识别的基团取代，并且其中R1是芳基或1-20个碳原子的烷基，其可以任选地被1个或多个卤素原子取代，并且其中T是在二氧杂环丁烷I的酶活化分解时能够发光的芳基或杂芳基环。还公开了包含二恶丁环化合物的方法和测定法。

8. 发明申请

US20130196325A1 In Situ Chemiluminescent Substrates and Assays 审中-公开
标题翻译：原位化学发光底物和测定
公开(公告)号：US20130196325A1
公开(公告)日：2013-08-01
申请号：US13808922
申请日：2011-07-06
申请人： Alison Sparks , Zhixian Wang , Melissa Gee
发明人： Alison Sparks , Zhixian Wang , Melissa Gee
IPC分类号： G01N33/58 , C12Q1/42 , C12Q1/68 , C12Q1/34
CPC分类号： G01N33/581 , C07D321/00 , C07F9/12 , C07F9/4056 , C07F9/6541 , C07F9/65512 , C07F9/65583 , C07H15/203 , C12Q1/34 , C12Q1/42 , C12Q1/6816
摘要： Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed.
摘要翻译：在水性或其他测定条件下原位生成化学发光酶底物的方法。还公开了使用底物产生光，检测和/或定量酶，抗原和/或核酸的方法。还披露了与这些方法有关的套件。

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