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1. 发明申请

WO2006104778A3 COMPOSITIONS AND METHODS FOR DETECTING PROTEASE ACTIVITY 审中-公开
公开(公告)号：WO2006104778A3
公开(公告)日：2006-10-05
申请号：PCT/US2006/010277
申请日：2006-03-21
申请人： ILLUMINA , INC. et al. , KOZLOV, Igor , MELNYK, Peter
发明人： KOZLOV, Igor , MELNYK, Peter
IPC分类号： C07B61/00 , C07H21/02 , C07H21/04 , C07K2/00 , C12N15/10 , C12Q1/68 , A61K38/00
摘要： The invention provides a method of determining activity of a protease. The method can include the steps of (a) providing a protease substrate including a protein moiety attached to a nucleic acid moiety and a ligand moiety; (b) contacting the protease substrate with a protease under conditions wherein the protease catalyzes cleavage of the protein moiety, thereby producing a proteolytic product wherein the nucleic acid moiety is separated from at least a portion of the protein moiety and the ligand moiety; (c) contacting the proteolytic product with a receptor under conditions wherein the ligand moiety binds to the receptor to form a complex; (d) separating the complex from the nucleic acid moiety, thereby forming a separation product including the nucleic acid moiety; (e) contacting the separation product with a probe nucleic acid under conditions wherein the nucleic acid moiety hybridizes to a complementary sequence of the probe; and (f) detecting hybridization of the separation product to the probe, thereby determining activity of the protease.

2. 发明申请

WO2007067923A3 COMPOSITIONS AND METHODS FOR DETECTING PHOSPHOMONOESTER 审中-公开
标题翻译：用于检测磷酸化合物的组合物和方法
公开(公告)号：WO2007067923A3
公开(公告)日：2007-11-22
申请号：PCT/US2006061681
申请日：2006-12-06
申请人： ILLUMINA INC , KOZLOV IGOR , MELNYK PETER , ZHAO CHANFENG
发明人： KOZLOV IGOR , MELNYK PETER , ZHAO CHANFENG
IPC分类号： G01N33/53 , C07K14/47
CPC分类号： G01N33/5308 , C07K1/1072 , C07K1/13
摘要： The invention provides a method of modifying a phosphomonoester moiety of a target compound. The method can include the steps of (a) providing a target compound having an electrophilic moiety and a phosphomonoester moiety; (b) contacting the target compound with a first carbodiimide compound under conditions for preferential addition of the first carbodiimide compound to the electrophilic moiety over the phosphomonoester moiety, thereby forming an electrophile-protected target compound; and (c) contacting the electrophile-protected target compound with a second carbodiimide compound and a nucleophilic compound under conditions for addition of the nucleophilic compound to the phosphomonoester.
摘要翻译：本发明提供了修饰目标化合物的磷酸单酯部分的方法。该方法可以包括以下步骤：（a）提供具有亲电子部分和磷酸单酯部分的目标化合物; （b）使所述目标化合物与第一碳化二亚胺化合物在优先向所述磷酸单酯部分上的所述亲电子部分加入所述第一碳二亚胺化合物的条件下接触，由此形成亲电子受保护的目标化合物; 和（c）在亲核化合物加入磷酸单酯的条件下，使亲电试剂保护的目标化合物与第二碳化二亚胺化合物和亲核化合物接触。

3. 发明申请

WO2007067923A2 COMPOSITIONS AND METHODS FOR DETECTING PHOSPHOMONOESTER 审中-公开
标题翻译：用于检测磷酸酯的组合物和方法
公开(公告)号：WO2007067923A2
公开(公告)日：2007-06-14
申请号：PCT/US2006/061681
申请日：2006-12-06
申请人： ILLUMINA, INC. , KOZLOV, Igor , MELNYK, Peter , ZHAO, Chanfeng
发明人： KOZLOV, Igor , MELNYK, Peter , ZHAO, Chanfeng
IPC分类号： G01N33/53 , C07K14/47
CPC分类号： G01N33/5308 , C07K1/1072 , C07K1/13
摘要： The invention provides a method of modifying a phosphomonoester moiety of a target compound. The method can include the steps of (a) providing a target compound having an electrophilic moiety and a phosphomonoester moiety; (b) contacting the target compound with a first carbodiimide compound under conditions for preferential addition of the first carbodiimide compound to the electrophilic moiety over the phosphomonoester moiety, thereby forming an electrophile-protected target compound; and (c) contacting the electrophile-protected target compound with a second carbodiimide compound and a nucleophilic compound under conditions for addition of the nucleophilic compound to the phosphomonoester.
摘要翻译：本发明提供了修饰目标化合物的磷酸单酯部分的方法。该方法可以包括以下步骤：（a）提供具有亲电子部分和磷酸酯部分的目标化合物; （b）在优先将第一碳二亚胺化合物添加到磷酸单酯部分上的亲电子部分的条件下，使目标化合物与第一碳二亚胺化合物接触，由此形成亲电试剂保护的目标化合物; 和（c）在向磷酸单酯添加亲核性化合物的条件下，将亲电试剂保护的目标化合物与第二碳二亚胺化合物和亲核化合物接触。

4. 发明申请

WO2006104778A2 COMPOSITIONS AND METHODS FOR DETECTING PROTEASE ACTIVITY 审中-公开
标题翻译：用于检测蛋白酶活性的组合物和方法
公开(公告)号：WO2006104778A2
公开(公告)日：2006-10-05
申请号：PCT/US2006010277
申请日：2006-03-21
申请人： ILLUMINA INC , KOZLOV IGOR , MELNYK PETER
发明人： KOZLOV IGOR , MELNYK PETER
IPC分类号： C12Q1/68
CPC分类号： C12Q1/37
摘要： The invention provides a method of determining activity of a protease. The method can include the steps of (a) providing a protease substrate including a protein moiety attached to a nucleic acid moiety and a ligand moiety; (b) contacting the protease substrate with a protease under conditions wherein the protease catalyzes cleavage of the protein moiety, thereby producing a proteolytic product wherein the nucleic acid moiety is separated from at least a portion of the protein moiety and the ligand moiety; (c) contacting the proteolytic product with a receptor under conditions wherein the ligand moiety binds to the receptor to form a complex; (d) separating the complex from the nucleic acid moiety, thereby forming a separation product including the nucleic acid moiety; (e) contacting the separation product with a probe nucleic acid under conditions wherein the nucleic acid moiety hybridizes to a complementary sequence of the probe; and (f) detecting hybridization of the separation product to the probe, thereby determining activity of the protease.
摘要翻译：本发明提供一种测定蛋白酶活性的方法。该方法可以包括以下步骤：（a）提供包含连接于核酸部分和配体部分的蛋白质部分的蛋白酶底物; （b）在所述蛋白酶底物与蛋白酶接触的条件下使所述蛋白酶底物接触，其中所述蛋白酶催化所述蛋白质部分的切割，由此产生蛋白水解产物，其中所述核酸部分与所述蛋白质部分和所述配体部分的至少一部分分离; （c）在其中配体部分结合受体以形成复合物的条件下使蛋白水解产物与受体接触; （d）从核酸部分分离复合物，从而形成包含核酸部分的分离产物; （e）在其中核酸部分与探针的互补序列杂交的条件下使分离产物与探针核酸接触; 和（f）检测分离产物与探针的杂交，从而确定蛋白酶的活性。

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