Compositions for the detection of protease in biological samples and
methods of use therefo
    1.
    发明授权
    Compositions for the detection of protease in biological samples and methods of use therefo 失效
    用于检测生物样品中蛋白酶的组合物及其使用方法

    公开(公告)号:US5714342A

    公开(公告)日:1998-02-03

    申请号:US549008

    申请日:1995-10-27

    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。

    Fluorogenic protease substrates based on dye-dimerization
    3.
    发明申请
    Fluorogenic protease substrates based on dye-dimerization 失效
    基于染料二聚化的荧光蛋白酶底物

    公开(公告)号:US20050089946A1

    公开(公告)日:2005-04-28

    申请号:US10930522

    申请日:2004-08-31

    摘要: A method of biological assay comprises the steps of providing an enzyme substrate comprising two fluorescence dye groups bound to a flexible peptide, the dye groups being of proximity sufficiently close so as to allow free energy attractions to draw the dyes together to essentially self-quench fluorescence of the dye groups, wherein self quenching of fluorescence of the dye groups is effected by dye dimerization or stacking, and enzymatically cleaving the peptide to release the fluorescence dye groups from dye dimerization or stacking, thereby producing an increase in fluorescence intensity. A protease substrate for use in the method of the invention is also disclosed. This invention finds use in detection and identification of microorganisms, sterilization assurance, pharmaceutical discovery, enzyme assays, immunoassays, and other biological assays.

    摘要翻译: 一种生物测定方法包括以下步骤:提供包含与柔性肽结合的两个荧光染料基团的酶底物,所述染料组的接近度足够接近,以便使自由能吸引力将染料一起吸收到基本上自熄灭荧光 的染料基团,其中染料基团的荧光自发淬灭通过染料二聚化或堆叠进行,并且酶切割肽以使荧光染料基团从染料二聚化或堆叠释放,从而产生荧光强度的增加。 还公开了用于本发明方法的蛋白酶底物。 本发明用于微生物的检测和鉴定,灭菌保证,药物发现,酶测定,免疫测定和其他生物测定。

    Assay reagent
    4.
    发明授权
    Assay reagent 失效
    分析试剂

    公开(公告)号:US5776720A

    公开(公告)日:1998-07-07

    申请号:US443776

    申请日:1995-05-18

    摘要: An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

    摘要翻译: 公开了一种用于测定代谢活性整个细胞内的酶的活性的测定化合物或其盐。 测定化合物包括离去基团和指示剂组。 离去基团选自包括氨基酸,肽,糖,硫酸盐,磷酸盐,酯,磷酸酯,核苷酸,多核苷酸,核酸,嘧啶,嘌呤,核苷,脂质及其混合物的组。 指示基团选自与离去基团连接时具有第一状态的化合物,当离去基团被酶从指示剂基团切割时的第二种状态。 优选地,指示剂化合物是这些化合物的罗丹明110,rhodol和荧光素以及类似物。 还公开了合成化合物的方法以及使用这些化合物测定酶活性的方法。

    Compositions for the detection of proteases in biological samples and
methods of use thereof
    5.
    发明授权
    Compositions for the detection of proteases in biological samples and methods of use thereof 失效
    用于检测生物样品中蛋白酶的组合物及其使用方法

    公开(公告)号:US5605809A

    公开(公告)日:1997-02-25

    申请号:US331383

    申请日:1994-10-28

    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。

    Fluorogenic protease substrates based on dye-dimerization
    6.
    发明授权
    Fluorogenic protease substrates based on dye-dimerization 失效
    基于染料二聚化的荧光蛋白酶底物

    公开(公告)号:US07256012B2

    公开(公告)日:2007-08-14

    申请号:US10930522

    申请日:2004-08-31

    摘要: A method of biological assay comprises the steps of providing an enzyme substrate comprising two fluorescence dye groups bound to a flexible peptide, the dye groups being of proximity sufficiently close so as to allow free energy attractions to draw the dyes together to essentially self-quench fluorescence of the dye groups, wherein self quenching of fluorescence of the dye groups is effected by dye dimerization or stacking, and enzymatically cleaving the peptide to release the fluorescence dye groups from dye dimerization or stacking, thereby producing an increase in fluorescence intensity. A protease substrate for use in the method of the invention is also disclosed. This invention finds use in detection and identification of microorganisms, sterilization assurance, pharmaceutical discovery, enzyme assays, immunoassays, and other biological assays.

    摘要翻译: 一种生物测定方法包括以下步骤:提供包含与柔性肽结合的两个荧光染料基团的酶底物,所述染料组的接近度足够接近,以便使自由能吸引力将染料一起吸收到基本上自熄灭荧光 的染料基团,其中染料基团的荧光自发淬灭通过染料二聚化或堆叠进行,并且酶切割肽以使荧光染料基团从染料二聚化或堆叠释放,从而产生荧光强度的增加。 还公开了用于本发明方法的蛋白酶底物。 本发明用于微生物的检测和鉴定,灭菌保证,药物发现,酶测定,免疫测定和其他生物测定。

    Fluorogenic protease substrates based on dye-dimerization
    7.
    发明授权
    Fluorogenic protease substrates based on dye-dimerization 失效
    基于染料二聚化的荧光蛋白酶底物

    公开(公告)号:US06787329B1

    公开(公告)日:2004-09-07

    申请号:US09448633

    申请日:1999-11-24

    IPC分类号: C12Q137

    摘要: A method of biological assay comprises the steps of providing an enzyme substrate comprising two fluorescence dye groups bound to a flexible peptide, the dye groups being of proximity sufficiently close so as to allow free energy attractions to draw the dyes together to essentially self-quench fluorescence of the dye groups, wherein self quenching of fluorescence of the dye groups is effected by dye dimerization or stacking, and enzymatically cleaving the peptide to release the fluorescence dye groups from dye dimerization or stacking, thereby producing an increase in fluorescence intensity. A protease substrate for use in the method of the invention is also disclosed. This invention finds use in detection and identification of microorganisms, sterilization assurance, pharmaceutical discovery, enzyme assays, immunoassays, and other biological assays.

    摘要翻译: 一种生物测定方法包括以下步骤:提供包含与柔性肽结合的两个荧光染料基团的酶底物,所述染料组的接近度足够接近,以便使自由能吸引力将染料一起吸收到基本上自熄灭荧光 的染料基团,其中染料基团的荧光自发淬灭通过染料二聚化或堆叠进行,并且酶切割肽以使荧光染料基团从染料二聚化或堆叠释放,从而产生荧光强度的增加。 还公开了用于本发明方法的蛋白酶底物。 本发明用于微生物的检测和鉴定,灭菌保证,药物发现,酶测定,免疫测定和其他生物测定。

    Assay reagent
    8.
    发明授权

    公开(公告)号:US5849513A

    公开(公告)日:1998-12-15

    申请号:US904400

    申请日:1997-07-31

    摘要: An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.