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    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
    1.
    发明授权
    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids 失效
    将寡核苷酸末端连接到用于捕获和检测核酸的聚丙烯酰胺固体支持物上

    公开(公告)号：US5478893A

    公开(公告)日：1995-12-26

    申请号：US102474

    申请日：1993-08-05

    申请人： Soumitra S. Ghosh ,  Eoin D. Fahy

    发明人： Soumitra S. Ghosh ,  Eoin D. Fahy

    IPC分类号： C07H21/00 ,  C12Q1/68 ,  C08F120/56 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C07H21/00

    摘要： Methods for covalent attachment of oligonucleotides to solid supports such that substantially all of the oligonucleotides are attached via their 5'-ends are provided. The solid supports with attached oligonucleotides are produced. Thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports. In a further aspect, bromoacetyl-derivatized oligonucleotides, and polyacrylamide supports with linked oligonucleotides produced by coupling bromoacetyl-derivatized oligonucleotides with thiol-derivatized polyacrylamide solid supports or by coupling thiol-derivatized oligonucleotides with bromoacetyl-derivatized polyacrylamide supports as well as methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats are provided.

    摘要翻译： 提供了将寡核苷酸共价连接到固体支持物的方法，使得基本上所有的寡核苷酸通过其5'-末端连接。 产生具有连接的寡核苷酸的固体支持物。 硫醇寡核苷酸连接到溴乙酰基衍生的聚丙烯酰胺载体上，或者相反地，溴乙酰基寡核苷酸固定在硫醇 - 聚丙烯酰胺载体上。 在另一方面，溴乙酰基衍生的寡核苷酸和具有通过将溴乙酰基衍生的寡核苷酸与硫醇衍生的聚丙烯酰胺固体支持物偶联或通过将硫醇衍生的寡核苷酸与溴乙酰基衍生的聚丙烯酰胺载体偶联产生的链接寡核苷酸的聚丙烯酰胺载体以及捕获 提供了通过直接捕获或以夹心杂交形式连接到聚丙烯酰胺固体支持物上的寡核苷酸的核酸。

    Sulfonamide-derivative compounds for tagging nucleic acid probes
    2.
    发明授权
    Sulfonamide-derivative compounds for tagging nucleic acid probes 失效
    用于标记核酸探针的磺酰胺衍生化合物

    公开(公告)号：US5210203A

    公开(公告)日：1993-05-11

    申请号：US879593

    申请日：1992-05-07

    申请人： Gary F. Musso ,  Soumitra Ghosh ,  Emil T. Kaiser

    发明人： Gary F. Musso ,  Soumitra Ghosh ,  Emil T. Kaiser

    IPC分类号： C07F7/18 ,  C07H21/00 ,  C12Q1/68

    CPC分类号： C07H21/00 ,  C07F7/1852 ,  C07F7/1856 ,  C12Q1/6816

    摘要： Nucleic acid hybridization probes are provided which comprise an N.sup.4 -(substituted amino)cytosine moiety, wherein the substituted amino group comprises a tag moiety, whereby the probe is detected. Methods of preparing probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in hybridization assays are also provided. Typical tag moieties employed with the probes of the invention are biotinyl, aminothiadiazole and fluorescein derivatives, connected to N.sup.4 -amino groups of modified cytosines of the probe through linker moieties. Probes tagged with biotin are typically detected by binding to the biotinyl moieties, through a streptavidin or avidin molecule, a reporter group which includes streptavidin or avidin and then detecting a signal due to the reporter group. Probes tagged with aminothiadiazole derivatives are typically detected by essentially the same method as those tagged with biotinyl but employing as reporter group one which binds to the derivative through a carbonic anhydrase molecule. Probes tagged with fluorescein derivatives are detected by a fluorescence spectroscopic method without binding of a reporter group to the tag.

    摘要翻译： 提供了包含N4-（取代氨基）胞嘧啶部分的核酸杂交探针，其中取代的氨基包括标签部分，由此检测探针。 还提供了制备本发明的探针的方法，在这些方法中使用的中间体以及在杂交测定中使用本发明的探针的方法。 与本发明的探针一起使用的典型标签部分是通过接头部分连接到探针的修饰胞嘧啶的N4-氨基的生物素，氨基噻二唑和荧光素衍生物。 用生物素标记的探针通常通过结合生物素部分，通过链霉抗生物素蛋白或抗生物素蛋白分子，包括链霉抗生物素蛋白或抗生物素蛋白的报道基团检测，然后检测由于报告基团引起的信号。 用氨基噻二唑衍生物标记的探针通常用与生物素标记的那些基本上相同的方法来检测，但采用作为通过碳酸酐酶分子与衍生物结合的报告基团。 用荧光素衍生物标记的探针通过荧光光谱法检测，而不将报道基团与标签结合。

    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
    3.
    发明授权
    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids 失效
    将寡核苷酸结合到聚丙烯酰胺固体支持物上用于捕获和检测核酸

    公开(公告)号：US5237016A

    公开(公告)日：1993-08-17

    申请号：US293893

    申请日：1989-01-05

    申请人： Soumitra S. Ghosh ,  Eoin D. Fahy

    发明人： Soumitra S. Ghosh ,  Eoin D. Fahy

    IPC分类号： C07H21/00 ,  C12Q1/68

    CPC分类号： C07H21/00

    摘要： This invention concerns methods and means for covalent attachment of oligonucleotides to solid supports substantially at their 5'-ends. According to the invention thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports.In a further aspect, this invention relates to bromoacetyl-oligonucleotides that may be immobilized on thiol-polyacrylamide solid supports, thiol-oligonucleotides immobilized on bromoacetyl-derivatized polyacrylamide supports as well as to methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats.

    摘要翻译： 本发明涉及将寡核苷酸共价连接到基本上在其5'端的固体支持物的方法和手段。 根据本发明，硫醇 - 寡核苷酸连接到溴乙酰基衍生的聚丙烯酰胺载体上，或者相反地，溴乙酰基寡核苷酸被固定在巯基 - 聚丙烯酰胺载体上。 在另一方面，本发明涉及可固定在硫醇 - 聚丙烯酰胺固体支持物上的溴乙酰基寡核苷酸，固定在溴乙酰基衍生的聚丙烯酰胺载体上的硫醇寡核苷酸以及通过与聚丙烯酰胺固体支持物连接的寡核苷酸捕获核酸的方法 ，通过直接捕获或夹心杂交形式。

    Carbonic anhydrase inhibitor-tagged nucleic acid probes
    5.
    发明授权
    Carbonic anhydrase inhibitor-tagged nucleic acid probes 失效
    碳酸酐酶抑制剂标记的核酸探针

    公开(公告)号：US4780405A

    公开(公告)日：1988-10-25

    申请号：US753176

    申请日：1985-07-09

    申请人： Emil T. Kaiser ,  Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl

    发明人： Emil T. Kaiser ,  Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl

    IPC分类号： G01N33/58 ,  C07F7/18 ,  C07H21/00 ,  C12N15/09 ,  C12Q1/68 ,  C12Q1/70 ,  G01N33/532 ,  G01N33/535

    CPC分类号： G01N33/535 ,  C07F7/1852 ,  C07F7/1856 ,  C07H21/00 ,  G01N33/532

    摘要： Nucleic acid hybridization probes are provided which comprise nucleoside bases or terminal nucleotide phosphates chemically linked to aromatic sulfonamide inhibitors of carbonic anhydrase. Methods of preparing probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in hybridization assays are also provided. A probe of the invention is detected by binding to it a reporter group, such as a homopolymer or heteropolymer of enzymes, which includes a carbonic anhydrase which binds to the inhibitor linked to the probe, and then detecting the bound reporter group, as by production of a fluorescent or colored product in a reaction catalyzed by an enzyme component of the reporter group. Also provided are enzyme immunoassays wherein detection of antibody is by a process which comprises a chemical reaction catalyzed by a carbonic anhydrase.

    摘要翻译： 提供核酸杂交探针，其包含与碳酸酐酶的芳族磺酰胺抑制剂化学连接的核苷碱基或末端核苷酸磷酸酯。 还提供了制备本发明的探针的方法，在这些方法中使用的中间体以及在杂交测定中使用本发明的探针的方法。 本发明的探针通过与其结合的报道基团如酶的均聚物或异源聚合物来检测，所述报告基团包括结合与探针连接的抑制剂的碳酸酐酶，然后通过生产检测结合的报道基团 的荧光或着色产物在由报道基团的酶组分催化的反应中。 还提供了酶免疫测定，其中抗体的检测是通过包含由碳酸酐酶催化的化学反应的方法。

    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
    6.
    发明授权
    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids 失效
    将寡核苷酸末端连接到用于捕获和检测核酸的聚丙烯酰胺固体支持物上

    公开(公告)号：US5663242A

    公开(公告)日：1997-09-02

    申请号：US414019

    申请日：1995-03-31

    申请人： Soumitra Shanker Ghosh ,  Eoin D. Fahy

    发明人： Soumitra Shanker Ghosh ,  Eoin D. Fahy

    IPC分类号： C07H21/00 ,  C12Q1/68 ,  C08F20/56 ,  C12P19/34 ,  C12Q1/70

    CPC分类号： C07H21/00

    摘要： Methods for covalent attachment of oligonucleotides to solid supports such that substantially all of the oligonucleotides are attached via their 5'-ends are provided. The solid supports with attached oligonucleotides are produced. Thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports. In a further aspect, bromoacetyl-derivatized oligonucleotides, and polyacrylamide supports with linked oligonucleotides produced by coupling bromoacetyl-derivatized oligonucleotides with thiol-derivatized polyacrylamide solid supports or by coupling thiol-derivatized oligonucleotides with bromoacetyl-derivatized polyacrylamide supports as well as methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats are provided.

    摘要翻译： 提供了将寡核苷酸共价连接到固体支持物的方法，使得基本上所有的寡核苷酸通过其5'-末端连接。 产生具有连接的寡核苷酸的固体支持物。 硫醇寡核苷酸连接到溴乙酰基衍生的聚丙烯酰胺载体上，或者相反地，溴乙酰基寡核苷酸固定在硫醇 - 聚丙烯酰胺载体上。 在另一方面，溴乙酰基衍生的寡核苷酸和具有通过将溴乙酰基衍生的寡核苷酸与硫醇衍生的聚丙烯酰胺固体支持物偶联或通过将硫醇衍生的寡核苷酸与溴乙酰基衍生的聚丙烯酰胺载体偶联产生的链接寡核苷酸的聚丙烯酰胺载体以及捕获 提供了通过直接捕获或以夹心杂交形式连接到聚丙烯酰胺固体支持物上的寡核苷酸的核酸。

    Compounds for tagging nucleic acid probes
    8.
    发明授权
    Compounds for tagging nucleic acid probes 失效
    用于标记核酸探针的化合物

    公开(公告)号：US4833251A

    公开(公告)日：1989-05-23

    申请号：US122496

    申请日：1987-11-16

    申请人： Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl ,  Emil T. Kaiser

    发明人： Gary F. Musso ,  Soumitra Ghosh ,  Leslie E. Orgel ,  Geoffrey M. Wahl ,  Emil T. Kaiser

    IPC分类号： C07F7/18 ,  C07H21/00 ,  C12Q1/68

    CPC分类号： C07H21/00 ,  C07F7/1852 ,  C07F7/1856 ,  C12Q1/6816

    摘要： Nucleic acid hybridization probes are provided which comprise an N.sup.4 -(substituted amino)cytosine moiety, wherein the substituted amino group comprises a tag moiety, whereby the probe is detected. Methods of preparing probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in hybridization assays are also provided. Typical tag moieties employed with the probes of the invention are biotinyl, aminothiadiazole and fluorescein derivatives, connected to N.sup.4 -amino groups of modified cytosines of the probe through linker moieties. Probes tagged with biotin are typically detected by binding to the biotinyl moieties, through a streptavidin or avidin molecule, a reporter group which includes streptavidin or avidin and then detecting a signal due to the reporter group. Probes tagged with aminothiadiazole derivatives are typically detected by essentially the same method as those tagged with biotinyl but employing as reporter group one which binds to the derivative through a carbonic anhydrase molecule. Probes tagged with fluorescein derivatives are detected by a fluorescence spectroscopic method without binding of a reporter group to the tag.

    摘要翻译： 提供了包含N4-（取代氨基）胞嘧啶部分的核酸杂交探针，其中取代的氨基包括标签部分，由此检测探针。 还提供了制备本发明的探针的方法，在这些方法中使用的中间体以及在杂交测定中使用本发明的探针的方法。 与本发明的探针一起使用的典型标签部分是通过接头部分连接到探针的修饰胞嘧啶的N4-氨基的生物素，氨基噻二唑和荧光素衍生物。 用生物素标记的探针通常通过结合生物素部分，通过链霉抗生物素蛋白或抗生物素蛋白分子，包括链霉抗生物素蛋白或抗生物素蛋白的报道基团检测，然后检测由于报告基团引起的信号。 用氨基噻二唑衍生物标记的探针通常用与生物素标记的那些基本上相同的方法来检测，但采用作为通过碳酸酐酶分子与衍生物结合的报告基团。 用荧光素衍生物标记的探针通过荧光光谱法检测，而不将报道基团与标签结合。