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1. 发明授权

US06183978B2 Luciferase assay, compositions and kits for use in connection therewith 有权
标题翻译：荧光素酶测定，用于与其结合使用的组合物和试剂盒
公开(公告)号：US06183978B2
公开(公告)日：2001-02-06
申请号：US09148782
申请日：1998-09-04
申请人： Irena Bronstein , Corinne E. M. Olesen , John C. Voyta , Yu-Xin Yan
发明人： Irena Bronstein , Corinne E. M. Olesen , John C. Voyta , Yu-Xin Yan
IPC分类号： C12Q166
CPC分类号： C12Q1/66
摘要： An assay for the presence of luciferase in a biological sample offers heightened sensitivity, signal intensity and persistence. The assay is sensitive down to 50 fg luciferase. The biological sample is combined with essential ingredients luciferin, ADP, myokinase and Mg++. The myokinase converts ADP to ATP, necessary for the luciferase reaction, and AMP, which retards the reaction kinetics. The resulting assay exhibits a persistent glow emission which makes it adaptable to automation.
摘要翻译：荧光素酶在生物样品中的存在检测提供了更高的灵敏度，信号强度和持续性。该测定法灵敏度低至50μg萤光素酶。生物样品与必需成分荧光素，ADP，肌激酶和Mg ++组合。肌动蛋白酶将ADP转化为ATP，这是荧光素酶反应所必需的，而AMP则延缓了反应动力学。所得测定显示持续的辉光发射，使其适应于自动化。

2. 发明授权

US5843681A Dioxetane compounds for the chemiluminescent detection of proteases, methods of use and kits therefore 失效
标题翻译：因此，用于化学发光检测蛋白酶的二氧杂环丁烷化合物，使用方法和试剂盒
公开(公告)号：US5843681A
公开(公告)日：1998-12-01
申请号：US728990
申请日：1996-10-11
申请人： Irena Bronstein , Brooks Edwards , Christopher Martin , Alison Sparks , John C. Voyta
发明人： Irena Bronstein , Brooks Edwards , Christopher Martin , Alison Sparks , John C. Voyta
IPC分类号： G01N33/573 , C07D321/00 , C07K5/072 , C07K5/08 , C07K5/09 , C07K5/10 , C12Q1/37 , G01N33/58
CPC分类号： C07K5/06095 , C07D321/00 , C07K5/0802 , C07K5/0817 , C07K5/1002 , C12Q1/37 , G01N33/582 , Y10S530/807
摘要： 1, 2-dioxetane compounds bearing a proteolytic enzyme specific amino acid or peptide are provided, which amino acid or peptide can be removed by action of the corresponding protease. When the amino acid or peptide is removed, the 1,2-dioxetane decomposes with chemiluminescence, the generation of light providing a rapid, ultra-sensitive and convenient means for detecting the presence of the protease in the sample being inspected. The amount of light generated, or degree of chemiluminescence, can be correlated with the amount of protease present. Immunoassays, as well as DNA hybridization and DNA probe assays are provided.
摘要翻译：提供了带有蛋白水解酶特异性氨基酸或肽的1,2-二氧杂环丁烷化合物，该氨基酸或肽可以通过相应的蛋白酶的作用除去。当去除氨基酸或肽时，1,2-二氧杂环丁烷以化学发光分解，产生光，提供快速，超灵敏和方便的手段，用于检测待检样品中蛋白酶的存在。产生的光量或化学发光度可以与存在的蛋白酶的量相关。提供免疫测定，以及DNA杂交和DNA探针测定。

3. 发明授权

US5591591A Dioxetane compounds for the chemiluminescent detection of proteases, methods of use and kits therefore 失效
标题翻译：用于化学发光检测蛋白酶的二氧杂环丁烷化合物，使用方法及其试剂盒
公开(公告)号：US5591591A
公开(公告)日：1997-01-07
申请号：US385788
申请日：1995-02-09
申请人： Irena Bronstein , Brooks Edwards , Christopher Martin , Alison Sparks , John C. Voyta
发明人： Irena Bronstein , Brooks Edwards , Christopher Martin , Alison Sparks , John C. Voyta
IPC分类号： G01N33/573 , C07D321/00 , C07K5/072 , C07K5/08 , C07K5/09 , C07K5/10 , C12Q1/37 , G01N33/58 , C07K5/06
CPC分类号： C07K5/06095 , C07D321/00 , C07K5/0802 , C07K5/0817 , C07K5/1002 , C12Q1/37 , G01N33/582 , Y10S530/807
摘要： 1,2-dioxetane compounds bearing a proteolytic enzyme specific amino acid or peptide are provided, which amino acid or peptide can be removed by action of the corresponding protease. When the amino acid or peptide is removed, the 1,2-dioxetane decomposes with chemiluminescence, the generation of light providing a rapid, ultra-sensitive and convenient means for detecting the presence of the protease in the sample being inspected. The amount of light generated, or degree of chemiluminescence, can be correlated with the amount of protease present. Immunoassays, as well as DNA hybridization and DNA probe assays are provided.
摘要翻译：提供了带有蛋白水解酶特异性氨基酸或肽的1,2-二氧环乙烷化合物，该氨基酸或肽可以通过相应的蛋白酶的作用除去。当去除氨基酸或肽时，1,2-二氧杂环丁烷以化学发光分解，产生光，提供快速，超灵敏和方便的手段，用于检测待检样品中蛋白酶的存在。产生的光量或化学发光度可以与存在的蛋白酶的量相关。提供免疫测定，以及DNA杂交和DNA探针测定。

4. 发明授权

US5783381A Chemiluminescent 1,2-dioxetanes 失效
公开(公告)号：US5783381A
公开(公告)日：1998-07-21
申请号：US545174
申请日：1995-10-19
申请人： Irena Bronstein , Brooks Edwards , Alison Sparks , John C. Voyta
发明人： Irena Bronstein , Brooks Edwards , Alison Sparks , John C. Voyta
IPC分类号： G01N31/00 , C07D305/06 , C07D321/00 , C07F7/18 , C09K11/07 , C12Q1/34 , G01N21/78 , G01N33/52 , C12Q1/00
CPC分类号： C12Q1/34 , C07D321/00 , C07F7/1856 , G01N33/52 , Y10S435/975
摘要： Dioxetanes that can be triggered by bases are provided which can be used to detect bases in organic solvents, aqueous preparations and detect gaseous bases in the atmosphere. The dioxetanes can be used to detect the release of bases from various labels, as a means to detect the presence of a base released by physical or natural processes, to calibrate light measuring apparatus and to determine the amount of reducing or oxidizing agent present in the base.

5. 发明授权

US06586196B1 Multiple enzyme assays 有权
标题翻译：多重酶测定
公开(公告)号：US06586196B1
公开(公告)日：2003-07-01
申请号：US09459982
申请日：1999-12-14
申请人： Irena Bronstein , Christopher Martin , Corinne Olesen , John Voyta , Yu-xin Yan
发明人： Irena Bronstein , Christopher Martin , Corinne Olesen , John Voyta , Yu-xin Yan
IPC分类号： C12Q142
CPC分类号： C12Q1/6897 , C12Q1/00 , C12Q1/34 , C12Q1/42 , C12Q1/66 , G01N2333/916 , G01N2333/924 , G01N2333/95
摘要： The present invention discloses multiple enzyme assays which measure the activity of at least one endogenous enzyme in a single aliquot and a method of measuring the activity of multiple enzymes in an aliquot of a cell extract, wherein at least one of the enzymes is an endogenous enzyme. In one embodiment of the invention the activity of a first enzyme is quantified by measuring the light signal produced by degradation of a first enzyme substrate by the first enzyme and the activity of the second enzyme is quantified by measuring the light signal produced by the degradation of a second substrate. In the method of the present invention, both quantifications are performed on the same aliquot of cell extract. Different embodiments of the present invention provide for the detection of more than one endogenous enzyme and for the detection of at least one reporter enzyme and at least one endogenous enzyme. The present invention also discloses kits for detecting the activity of multiple enzymes.
摘要翻译：本发明公开了测量单一等分试样中至少一种内源性酶的活性的多种酶测定法和测量细胞提取物等分试样中多种酶的活性的方法，其中至少一种酶是内源性酶。在本发明的一个实施方案中，通过测量由第一酶降解第一酶底物产生的光信号来定量第一酶的活性，并且通过测量由第一酶降解产生的光信号来量化第二酶的活性第二基板。在本发明的方法中，两种定量在相同等分的细胞提取物上进行。本发明的不同实施方案提供了多于一种内源性酶的检测和用于检测至少一种报道酶和至少一种内源性酶。本发明还公开了用于检测多种酶的活性的试剂盒。

6. 发明授权

US6132956A Chemiluminescent 1,2-dioxetanes 失效
标题翻译：化学发光的1,2-二氧杂环丁烷
公开(公告)号：US6132956A
公开(公告)日：2000-10-17
申请号：US964552
申请日：1997-11-05
申请人： Irena Bronstein , Brooks Edwards , Alison Sparks , John C. Voyta
发明人： Irena Bronstein , Brooks Edwards , Alison Sparks , John C. Voyta
IPC分类号： G01N31/00 , C07D305/06 , C07D321/00 , C07F7/18 , C09K11/07 , C12Q1/34 , G01N21/78 , G01N33/52 , C12Q1/00
CPC分类号： C12Q1/34 , C07D321/00 , C07F7/1856 , G01N33/52 , Y10S435/975
摘要： Dioxetanes that can be triggered by bases are provided which can be used to detect bases in organic solvents, aqueous preparations and detect gaseous bases in the atmosphere. The dioxetanes can be used to detect the release of bases from various labels, as a means to detect the presence of a base released by physical or natural processes, to calibrate light measuring apparatus and to determine the amount of reducing or oxidizing agent present in the base.
摘要翻译：提供可由碱触发的二氧杂环丁烷，可用于检测有机溶剂，含水制剂中的碱，并检测大气中的气态碱。二氧杂环丁烷可用于检测各种标记物中碱的释放，作为检测通过物理或天然过程释放的碱的存在的手段，校准光测量装置并确定存在于基础。

7. 发明授权

US5871938A Dioxetane compounds for the chemiluminescent detection of proteases, methods of use and kits therefore 失效
标题翻译：因此，用于化学发光检测蛋白酶的二氧杂环丁烷化合物，使用方法和试剂盒
公开(公告)号：US5871938A
公开(公告)日：1999-02-16
申请号：US935066
申请日：1997-09-22
申请人： Irena Bronstein , Brooks Edwards , Christopher Martin , Alison Sparks , John C. Voyta
发明人： Irena Bronstein , Brooks Edwards , Christopher Martin , Alison Sparks , John C. Voyta
IPC分类号： G01N33/573 , C07D321/00 , C07K5/072 , C07K5/08 , C07K5/09 , C07K5/10 , C12Q1/37 , G01N33/58 , C07K5/06
CPC分类号： C07K5/06095 , C07D321/00 , C07K5/0802 , C07K5/0817 , C07K5/1002 , C12Q1/37 , G01N33/582 , Y10S530/807
摘要： 1,2-dioxetane compounds bearing a proteolytic enzyme specific amino acid or peptide are provided, which amino acid or peptide can be removed by action of the corresponding protease. When the amino acid or peptide is removed, the 1,2-dioxetane decomposes with chemiluminescence, the generation of light providing a rapid, ultra-sensitive and convenient means for detecting the presence of the protease in the sample being inspected. The amount of light generated, or degree of chemiluminescence, can be correlated with the amount of protease present. Immunoassays, as well as DNA hybridization and DNA probe assays are provided.
摘要翻译：提供了带有蛋白水解酶特异性氨基酸或肽的1,2-二氧环乙烷化合物，该氨基酸或肽可以通过相应的蛋白酶的作用除去。当去除氨基酸或肽时，1,2-二氧杂环丁烷以化学发光分解，产生光，提供快速，超灵敏和方便的手段，用于检测待检样品中蛋白酶的存在。产生的光量或化学发光度可以与存在的蛋白酶的量相关。提供免疫测定，以及DNA杂交和DNA探针测定。

8. 发明授权

US06852548B2 Benzothiazole dioxetanes 有权
标题翻译：苯并噻唑二氧杂环丁烷
公开(公告)号：US06852548B2
公开(公告)日：2005-02-08
申请号：US09945661
申请日：2001-09-05
申请人： Brooks Edwards , Irena Bronstein
发明人： Brooks Edwards , Irena Bronstein
IPC分类号： G01N21/76 , C07D277/62 , C07D417/04 , C07D493/04 , C07F9/655 , C07F9/6558 , C12Q1/42 , G01N21/78 , G01N33/58 , G01N33/533 , G01N33/532
CPC分类号： C07D277/62 , C07D417/04 , C07D493/04 , C07F9/65512 , C07F9/65586 , G01N33/582 , Y10S530/807
摘要： Chemiluminescent 1,2-dioxetane compounds capable of producing light energy when decomposed, substantially stable at room temperature, represented by the formulas I or II: wherein T is:
摘要翻译：化学发光的1,2-二氧杂环丁烷化合物当分解时能够产生光能，在室温下基本稳定，由式I或II表示：其中T为：

9. 发明授权

US6140495A Chemiluminescent 1,2-dioxetanes 失效
标题翻译：化学发光的1,2-二氧杂环丁烷
公开(公告)号：US6140495A
公开(公告)日：2000-10-31
申请号：US296539
申请日：1999-04-22
申请人： Irena Bronstein , Brooks Edwards , Alison Sparks
发明人： Irena Bronstein , Brooks Edwards , Alison Sparks
IPC分类号： C07D321/00 , C07D327/06 , C07F9/02 , C07F9/06 , C07F9/28 , C07H19/04
CPC分类号： C07D321/00
摘要： Spiroadamantyl dioxetanes bearing an alkoxy substituent, and an aromatic substituent of phenyl or naphthyl on the dioxetane ring can be activated to chemiluminesce if the aromatic substituent bears a moiety designated OX, wherein the X is cleaved by an enzyme with which the dioxetane is permitted to come in contact with. The T.sub.1/2 kinetics of the chemiluminescent reaction, as well as the signal intensity, or quantum yield of the chemiluminescent reaction, can be altered by selection of an electron-withdrawing or an electron-donating group Z, at positions on the aromatic substituent other than those adjacent the point of attachment to the dioxetane. Signal strength can further be enhanced by recognized chemiluminescent enhancers.
摘要翻译：具有烷氧基取代基的双金刚烷基二氧杂环丁烷和二氧杂环丁烷环上苯基或萘基的芳族取代基可以被活化成化学发光，如果芳族取代基具有指定为OX的部分，其中X被允许二氧杂环丁烷的酶切割在与接触。化学发光反应的T + E，1/2 + EE动力学以及化学发光反应的信号强度或量子产率可以通过选择吸电子基团或给电子基团Z来改变，在芳族取代基上除了与二氧杂环丁烷的连接点相邻的位置之外的位置。信号强度可以通过认可的化学发光增强剂进一步增强。

10. 发明授权

US5800999A Dioxetane-precursor-labeled probes and detection assays employing the same 失效
标题翻译：二氧杂环丁烷 - 前体标记的探针和使用其的检测测定
公开(公告)号：US5800999A
公开(公告)日：1998-09-01
申请号：US767479
申请日：1996-12-16
申请人： Irena Bronstein , Brooks Edwards , Christopher Martin , John Voyta
发明人： Irena Bronstein , Brooks Edwards , Christopher Martin , John Voyta
IPC分类号： G01N33/532 , C07D321/00 , C07H21/00 , C07H21/02 , C07H21/04 , C12N15/09 , C12Q1/68 , C12N15/11
CPC分类号： C07H21/00 , C07D321/00 , C12Q1/6816 , C12Q1/686 , Y02P20/55
摘要： Probes labeled with 1,2-dioxetane precursors can be employed in a variety of assays. The probes may be nucleic acid, peptide nucleic acid, proteins including enzyme, antibody or antigen, steroid, carbohydrate, drug or non-drug hapten. The probe is provided with a 1,2-dioxetane precursor bound thereto, generally either covalently, or a strong ligand bond. The dioxetane precursor moiety is converted to a bound 1,2-dioxetane by exposure to singlet oxygen. These dioxetane (labels) either spontaneously decompose, or are induced to decompose by an appropriate trigger to release light. The trigger may be a change in pH temperature, or an agent which removes a protective group. Assay formats in which these 1,2-dioxetane labeled probes and referents may be used to include hybridization assays, immuno assays, gel-based assays and Capillary Zone Electrophoresis.
摘要翻译：用1,2-二氧杂环丁烷前体标记的探针可用于多种测定。探针可以是核酸，肽核酸，蛋白质，包括酶，抗体或抗原，类固醇，碳水化合物，药物或非药物半抗原。探针具有与其结合的1,2-二氧杂环丁烷前体，通常共价或强配体键。通过暴露于单线态氧将二氧杂环丁烷前体部分转化为结合的1,2-二氧环乙烷。这些二氧杂环丁烷（标记）会自发分解，或被适当的触发剂诱导分解以释放光。触发可能是pH温度的变化，或者是除去保护基团的试剂。这些1,2-二氧杂环丁烷标记的探针和指示物可用于包括杂交测定，免疫测定，基于凝胶的测定和毛细管区域电泳的测定形式。

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