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公开(公告)号：US08445191B2
公开(公告)日：2013-05-21
申请号：US13068144
申请日：2011-05-02
CPC分类号： G01N33/581 , A61K47/6883 , A61K49/0058 , A61K49/0067 , C07K17/08 , G01N33/532 , G01N33/54353 , G01N33/544 , G01N33/545 , G01N33/56983 , G01N33/57492 , G01N33/582 , G01N33/587 , G01N33/588
摘要： Certain disclosed embodiments of the present invention concern the synthesis, derivatization, conjugation to immunoglobulins and signal amplification based on discrete, relatively short polymers having plural reactive functional groups that react with plural molecules of interest. Reactive functional groups, such as hydrazides, may be derivatized with a variety of detectable labels, particularly haptens. The remaining reactive functional groups may be conjugated directly to a specific binding molecule, such as to the oxidized carbohydrate of the Fc region of the antibody. Disclosed conjugates display large signal amplification as compared to those based on molecules derivatized with single haptens, and are useful for assay methods, particularly multiplexed assays.
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公开(公告)号：US08658389B2
公开(公告)日：2014-02-25
申请号：US12381638
申请日：2009-03-13
申请人： Christopher Bieniarz , Jennifer Wong , Mark Lefever , Jerome W. Kosmeder, II , Julia Ashworth-Sharpe , Casey A. Kernag
发明人： Christopher Bieniarz , Jennifer Wong , Mark Lefever , Jerome W. Kosmeder, II , Julia Ashworth-Sharpe , Casey A. Kernag
IPC分类号： G01N33/535 , G01N33/53 , G01N33/532 , G01N21/76
CPC分类号： G01N33/535 , A61K47/65 , A61K47/6815 , C07K16/42 , C07K16/44 , C07K2317/76 , C12N9/0065 , C12N9/16 , C12N9/96
摘要： Antibody/signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification.
摘要翻译： 公开了抗体/信号产生部分缀合物，其包括通过异双功能聚亚烷基二醇连接体与产生信号的部分共价连接的抗体。 所公开的缀合物在组织切片和细胞学样品的免疫组织化学和原位杂交测定中显示出异常的信号产生。 在一个实施方案中，用半抗原标记的探针对核酸序列的酶 - 金相检测可以使用所公开的缀合物作为一级抗体而无需扩增来完成。
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公开(公告)号：US20090176253A1
公开(公告)日：2009-07-09
申请号：US12381638
申请日：2009-03-13
申请人： Christopher Bieniarz , Jennifer Wong , Mark Lefever , Jerome W. Kosmeder , Julia Ashworth-Sharpe , Casey A. Kernag
发明人： Christopher Bieniarz , Jennifer Wong , Mark Lefever , Jerome W. Kosmeder , Julia Ashworth-Sharpe , Casey A. Kernag
IPC分类号： G01N33/542 , G01N33/53
CPC分类号： G01N33/535 , A61K47/65 , A61K47/6815 , C07K16/42 , C07K16/44 , C07K2317/76 , C12N9/0065 , C12N9/16 , C12N9/96
摘要： Antibody/signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification.
摘要翻译： 公开了抗体/信号产生部分缀合物，其包括通过异双功能聚亚烷基二醇连接体与产生信号的部分共价连接的抗体。 所公开的缀合物在组织切片和细胞学样品的免疫组织化学和原位杂交测定中显示出异常的信号产生。 在一个实施方案中，用半抗原标记的探针对核酸序列的酶 - 金相检测可以使用所公开的缀合物作为一级抗体而无需扩增来完成。
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公开(公告)号：US08686122B2
公开(公告)日：2014-04-01
申请号：US12687564
申请日：2010-01-14
申请人： Christopher Bieniarz , Julia Ashworth-Sharpe , Casey A. Kernag , Jerome W. Kosmeder , Mark Lefever
发明人： Christopher Bieniarz , Julia Ashworth-Sharpe , Casey A. Kernag , Jerome W. Kosmeder , Mark Lefever
IPC分类号： C07K1/13 , C07K1/10 , G01N33/532
CPC分类号： G01N33/581 , A61K39/44 , A61K47/6801 , A61K47/6803 , A61K47/6815 , C07C319/02 , C07C323/22 , C07C323/60 , C07C327/28 , C07K16/00 , C12N9/16 , C12N9/96 , C12Y301/03001 , G01N33/532 , G01N33/535 , C07C323/12
摘要： A method is disclosed for making a conjugate of two molecules using a hydrazide thiol linker. In a particular working embodiment, an Fc-specific antibody-enzyme conjugate is made using the method and demonstrated to provide exceptional staining sensitivity and specificity in immunohistochemical and in situ hybridization assays.
摘要翻译： 公开了使用酰肼硫醇连接体制备两个分子的缀合物的方法。 在特定的工作实施方案中，使用该方法制备Fc特异性抗体 - 酶缀合物，并证实其在免疫组织化学和原位杂交测定中提供特殊的染色敏感性和特异性。
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公开(公告)号：US20100136652A1
公开(公告)日：2010-06-03
申请号：US12687564
申请日：2010-01-14
申请人： Christopher Bieniarz , Julia Ashworth-Sharpe , Casey A. Kernag , Jerome W. Kosmeder , Mark Lefever
发明人： Christopher Bieniarz , Julia Ashworth-Sharpe , Casey A. Kernag , Jerome W. Kosmeder , Mark Lefever
IPC分类号： C12N9/08 , C07C241/00 , C07K16/00 , C07D249/04 , C12N9/00 , C12N9/18 , C07K16/46
CPC分类号： G01N33/581 , A61K39/44 , A61K47/6801 , A61K47/6803 , A61K47/6815 , C07C319/02 , C07C323/22 , C07C323/60 , C07C327/28 , C07K16/00 , C12N9/16 , C12N9/96 , C12Y301/03001 , G01N33/532 , G01N33/535 , C07C323/12
摘要： A method is disclosed for making a conjugate of two molecules using a hydrazide thiol linker. In a particular working embodiment, an Fc-specific antibody-enzyme conjugate is made using the method and demonstrated to provide exceptional staining sensitivity and specificity in immunohistochemical and in situ hybridization assays.
摘要翻译： 公开了使用酰肼硫醇连接体制备两个分子的缀合物的方法。 在特定的工作实施方案中，使用该方法制备Fc特异性抗体 - 酶缀合物，并证实其在免疫组织化学和原位杂交测定中提供特殊的染色敏感性和特异性。
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6.发明授权Enzyme-catalyzed metal deposition for the enhanced in situ detection of immunohistochemical epitopes and nucleic acid sequences 有权酶催化的金属沉积用于增强免疫组织化学表位和核酸序列的原位检测公开(公告)号：US07632652B2
公开(公告)日：2009-12-15
申请号：US10877919
申请日：2004-06-24
IPC分类号： C12Q1/42 , C12Q1/44 , C12Q1/54 , C12Q1/68 , G01N33/53 , G01N33/543 , G01N33/567
CPC分类号： G01N33/581 , C07F9/093 , C07F9/65517 , C07F9/65522 , C07H15/20 , C07H15/24 , C07H15/26
摘要： The invention is directed to novel compositions of matter and methods of detecting in situ an immunohistochemical epitope or nucleic acid sequence of interest in a biological sample comprising binding an enzyme-labeled conjugate molecule to the epitope or sequence of interest in the presence of a redox-inactive reductive species and a soluble metal ion, thereby facilitating the reduction of the metal ion to a metal atom at or about the point where the enzyme is anchored. Novel phosphate derivatives of reducing agents are described that when exposed to a phosphatase are activated to their reducing form, thereby reducing metal ions to insoluble metal.
摘要翻译： 本发明涉及物质的新组合物和原位检测生物样品中感兴趣的免疫组织化学表位或核酸序列的方法，包括在氧化还原反应的存在下将酶标记的缀合物分子与感兴趣的表位或序列结合， 无活性还原物质和可溶性金属离子，从而有助于金属离子在酶被锚定的位置处或附近还原成金属原子。 描述了还原剂的新型磷酸酯衍生物，当暴露于磷酸酶被活化成其还原形式时，从而将金属离子还原成不溶性金属。
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公开(公告)号：US20110245098A1
公开(公告)日：2011-10-06
申请号：US13066980
申请日：2011-04-29
CPC分类号： G01N33/581 , A61K47/6883 , A61K49/0058 , A61K49/0067 , C07K17/08 , G01N33/532 , G01N33/54353 , G01N33/544 , G01N33/545 , G01N33/56983 , G01N33/57492 , G01N33/582 , G01N33/587 , G01N33/588
摘要： Certain disclosed embodiments of the present invention concern the synthesis, derivatization, conjugation to immunoglobulins and signal amplification based on discrete, relatively short polymers having plural reactive functional groups that react with plural molecules of interest. Reactive functional groups, such as hydrazides, may be derivatized with a variety of detectable labels, particularly haptens. The remaining reactive functional groups may be conjugated directly to a specific binding molecule, such as to the oxidized carbohydrate of the Fc region of the antibody. Disclosed conjugates display large signal amplification as compared to those based on molecules derivatized with single haptens, and are useful for assay methods, particularly multiplexed assays.
摘要翻译： 本发明的某些公开的实施方案涉及到具有多个与多个感兴趣的分子反应的反应性官能团的离散的，较短的聚合物的合成，衍生化，免疫球蛋白的缀合和信号扩增。 反应性官能团，例如酰肼，可用各种可检测的标记，特别是半抗原衍生化。 剩余的反应性官能团可以直接与特异性结合分子结合，例如与抗体Fc区的氧化碳水化合物结合。 与基于用单个半抗原衍生的分子相比，所公开的缀合物显示出大的信号扩增，并且可用于测定方法，特别是多重测定。
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公开(公告)号：US07985557B2
公开(公告)日：2011-07-26
申请号：US12154472
申请日：2008-05-22
CPC分类号： G01N33/581 , A61K47/6883 , A61K49/0058 , A61K49/0067 , C07K17/08 , G01N33/532 , G01N33/54353 , G01N33/544 , G01N33/545 , G01N33/56983 , G01N33/57492 , G01N33/582 , G01N33/587 , G01N33/588
摘要： Certain disclosed embodiments of the present invention concern the synthesis, derivatization, conjugation to immunoglobulins and signal amplification based on discrete, relatively short polymers having plural reactive functional groups that react with plural molecules of interest. Reactive functional groups, such as hydrazides, may be derivatized with a variety of detectable labels, particularly haptens. The remaining reactive functional groups may be conjugated directly to a specific binding molecule, such as to the oxidized carbohydrate of the Fc region of the antibody. Disclosed conjugates display large signal amplification as compared to those based on molecules derivatized with single haptens, and are useful for assay methods, particularly multiplexed assays.
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公开(公告)号：US20110244545A1
公开(公告)日：2011-10-06
申请号：US13068145
申请日：2011-05-02
CPC分类号： G01N33/581 , A61K47/6883 , A61K49/0058 , A61K49/0067 , C07K17/08 , G01N33/532 , G01N33/54353 , G01N33/544 , G01N33/545 , G01N33/56983 , G01N33/57492 , G01N33/582 , G01N33/587 , G01N33/588
摘要： Certain disclosed embodiments of the present invention concern the synthesis, derivatization, conjugation to immunoglobulins and signal amplification based on discrete, relatively short polymers having plural reactive functional groups that react with plural molecules of interest. Reactive functional groups, such as hydrazides, may be derivatized with a variety of detectable labels, particularly haptens. The remaining reactive functional groups may be conjugated directly to a specific binding molecule, such as to the oxidized carbohydrate of the Fc region of the antibody. Disclosed conjugates display large signal amplification as compared to those based on molecules derivatized with single haptens, and are useful for assay methods, particularly multiplexed assays.
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10.发明申请公开(公告)号：US20100297725A1
公开(公告)日：2010-11-25
申请号：US12660744
申请日：2010-03-02
申请人： Jerome W. Kosmeder , Mark Lefever , Donald Johnson , Michael Farrell , Zhanna Zhilina , Christopher Bieniarz
发明人： Jerome W. Kosmeder , Mark Lefever , Donald Johnson , Michael Farrell , Zhanna Zhilina , Christopher Bieniarz
CPC分类号： G01N33/5308 , C07D495/04 , C12Q1/6804 , C12Q1/6841 , C12Q1/6886 , G01N33/532 , G01N33/533 , G01N33/58 , G01N33/582 , G01N33/583 , Y10S435/961 , Y10S530/807
摘要： A method for performing a multiplexed diagnostic assay, such as for two or more different targets in a sample, is described. One embodiment comprised contacting the sample with two or more specific binding moieties that bind specifically to two or more different targets. The two or more specific binding moieties are conjugated to different haptens, and at least one of the haptens is an oxazole, a pyrazole, a thiazole, a nitroaryl compound other than dinitrophenyl, a benzofurazan, a triterpene, a urea, a thiourea, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azo aryl, or a benzodiazepine. The sample is contacted with two or more different anti-hapten antibodies that can be detected separately. The two or more different anti-hapten antibodies may be conjugated to different detectable labels.
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