公开(公告)号：US20090325168A1

公开(公告)日：2009-12-31

申请号：US12422127

申请日：2009-04-10

申请人： Beverly S. Packard ,  Akira Komoriya

发明人： Beverly S. Packard ,  Akira Komoriya

IPC分类号： C12Q1/68 ,  C12Q1/02 ,  C12N5/06

CPC分类号： C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  C12Q1/44 ,  G01N33/533 ,  G01N33/542 ,  G01N2333/918 ,  G01N2333/96425 ,  G01N2333/96469

摘要： The present invention provides for novel reagents whose fluorescence changes upon cleavage or a change in conformation of a backbone. The reagents comprise a backbone (e.g. nucleic acid, polypeptide, etc.) joining two fluorophores of the same species whereby the fluorophores form an H-dimer resulting in quenching of the fluorescence of the fluorophores. When the backbone is cleaved or changes conformation, the fluorophores are separated, no longer forming an H-type dimer, and are de-quenched thereby providing a detectable signal. The use of a single fluorophore rather than an “acceptor-donor” fluoresecence resonance energy transfer system offers synthesis and performance advantages.

摘要翻译： 本发明提供了新的试剂，其荧光在裂解时发生变化或骨架的构象变化。 试剂包含连接相同物质的两个荧光团的骨架（例如核酸，多肽等），由此荧光团形成H-二聚体，导致荧光团的荧光猝灭。 当主链被切割或改变构象时，荧光团被分离，不再形成H型二聚体，并且被淬灭，从而提供可检测的信号。 使用单个荧光团而不是“受体 - 供体”荧光共振能量转移系统提供综合和性能优势。

公开(公告)号：US6037137A

公开(公告)日：2000-03-14

申请号：US802981

申请日：1997-02-20

申请人： Akira Komoriya ,  Beverly S. Packard

发明人： Akira Komoriya ,  Beverly S. Packard

IPC分类号： C12N15/09 ,  C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  C12Q1/68 ,  G01N33/542 ,  A61K38/00 ,  G01N21/00 ,  G01N21/76

CPC分类号： C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/542 ,  G01N2333/96469

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone each end of which is conjugated to a fluorophore. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包括特征性折叠的肽主链，其每个末端与荧光团缀合。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于其可见光波长的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。

公开(公告)号：US06936687B1

公开(公告)日：2005-08-30

申请号：US09394019

申请日：1999-09-10

申请人： Akira Komoriya ,  Beverly S. Packard

发明人： Akira Komoriya ,  Beverly S. Packard

IPC分类号： C12N15/09 ,  C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  C12Q1/68 ,  G01N33/542 ,  A61K38/00

CPC分类号： C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/542 ,  G01N2333/96469

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone each end of which is conjugated to a fluorophore. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包括特征性折叠的肽主链，其每个末端与荧光团缀合。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于其可见光波长的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。

公开(公告)号：US07879574B2

公开(公告)日：2011-02-01

申请号：US11941766

申请日：2007-11-16

申请人： Beverly S. Packard ,  Akira Komoriya

发明人： Beverly S. Packard ,  Akira Komoriya

IPC分类号： C07K7/00 ,  C12Q1/37

CPC分类号： C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/542 ,  G01N2333/96469

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone conjugated to two fluorophores such that the fluorophores are located opposite sides of a cleavage site. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high specificity and their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包含与两个荧光团缀合的特征性折叠的肽主链，使得荧光团位于切割位点的相对侧。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于它们的高特异性和在可见波长中的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中的蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。

公开(公告)号：US5605809A

公开(公告)日：1997-02-25

申请号：US331383

申请日：1994-10-28

申请人： Akira Komoriya ,  Beverly S. Packard

发明人： Akira Komoriya ,  Beverly S. Packard

IPC分类号： G01N33/566 ,  A61K38/00 ,  C07K5/10 ,  C07K7/06 ,  C07K7/08 ,  C07K14/81 ,  C09K11/07 ,  C12Q1/37 ,  C12Q1/00 ,  G01N33/00

CPC分类号： C07K7/06 ,  C07K14/811 ,  C07K7/08 ,  C12Q1/37 ,  A61K38/00 ,  C12Q2334/20 ,  C12Q2337/40 ,  Y10S435/968 ,  Y10S436/80

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone each end of which is conjugated to a fluorophore. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包括特征性折叠的肽主链，其每个末端与荧光团缀合。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于其可见光波长的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。

公开(公告)号：US07312302B2

公开(公告)日：2007-12-25

申请号：US09874350

申请日：2001-06-04

申请人： Beverly S. Packard ,  Akira Komoriya

发明人： Beverly S. Packard ,  Akira Komoriya

IPC分类号： C12Q1/37 ,  C07K7/00

CPC分类号： C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/533 ,  G01N33/542 ,  G01N2333/96425 ,  G01N2333/96469

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone conjugated to two fluorophores such that the fluorophores are located opposite sides of a cleavage site. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high specificity and their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包含与两个荧光团缀合的特征性折叠的肽主链，使得荧光团位于切割位点的相对侧。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于它们的高特异性和在可见波长中的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中的蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。

公开(公告)号：US20080199898A1

公开(公告)日：2008-08-21

申请号：US11941766

申请日：2007-11-16

申请人： Beverly S. Packard ,  Akira Komoriya

发明人： Beverly S. Packard ,  Akira Komoriya

IPC分类号： C12Q1/37 ,  C12N5/00

CPC分类号： C07K7/06 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/542 ,  G01N2333/96469

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone conjugated to two fluorophores such that the fluorophores are located opposite sides of a cleavage site. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high specificity and their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包含与两个荧光团缀合的特征性折叠的肽主链，使得荧光团位于切割位点的相对侧。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于它们的高特异性和在可见波长中的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中的蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。

公开(公告)号：US5714342A

公开(公告)日：1998-02-03

申请号：US549008

申请日：1995-10-27

申请人： Akira Komoriya ,  Beverly S. Packard

发明人： Akira Komoriya ,  Beverly S. Packard

IPC分类号： G01N33/566 ,  A61K38/00 ,  C07K5/10 ,  C07K7/06 ,  C07K7/08 ,  C07K14/81 ,  C09K11/07 ,  C12Q1/37 ,  C07D311/88 ,  C09K11/06 ,  C12Q1/00

CPC分类号： C07K7/06 ,  C07K14/811 ,  C07K7/08 ,  C12Q1/37 ,  A61K38/00 ,  C12Q2334/20 ,  C12Q2337/40 ,  Y10S435/968 ,  Y10S436/80

摘要： The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone each end of which is conjugated to a fluorophore. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

摘要翻译： 本发明提供了在特定蛋白酶存在下荧光增加的新型试剂。 试剂包括特征性折叠的肽主链，其每个末端与荧光团缀合。 当折叠的肽被切割时，通过用蛋白酶消化，荧光团提供可见波长的高强度荧光信号。 由于其可见光波长的高荧光信号，这些蛋白酶指示剂特别适用于检测生物样品中蛋白酶活性，特别是在冷冻组织切片中。 因此，本发明还提供了在冷冻切片中原位检测蛋白酶活性的方法。