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1. 发明申请

US20100151489A1 Method for quantifying biomolecules conjugated to a nanoparticle 审中-公开
公开(公告)号：US20100151489A1
公开(公告)日：2010-06-17
申请号：US12658092
申请日：2010-02-01
申请人： Xiao-Bo Chen , Christopher Bieniarz , Michael Farrell
发明人： Xiao-Bo Chen , Christopher Bieniarz , Michael Farrell
IPC分类号： G01N33/53
CPC分类号： C12Q1/37 , B82Y15/00 , G01N33/54313 , G01N33/582 , Y10T436/143333 , Y10T436/17 , Y10T436/25125
摘要： Disclosed embodiments concern quantifying a biomolecule conjugated to a nanoparticle. Quantifying typically comprises determining the number of biomolecules per nanoparticle. Any suitable biomolecule can be used, including but not limited to, amino acids, peptides, proteins, haptens, nucleic acids, oligonucleotides, DNA, RNA, and combinations thereof. A single type of biomolecule may be conjugated to the nanoparticle, more than one biomolecule of a particular class may be conjugated to the nanoparticle, or two or more classes of biomolecules may be conjugated to the nanoparticle. Certain disclosed embodiments comprise enzymatically or chemically digesting a biomolecule conjugated to the nanoparticle, or displacing a biomolecule using ligand-exchange chemistry. Where biomolecule concentrations are determined, any technique suitable for determining biomolecule concentration can be used, such as spectrophotometric techniques, including measuring tryptophan fluorescence and using a standard fluorescence intensity versus biomolecule concentration curve.

2. 发明申请

US20080274463A1 Method for quantifying biomolecules conjugated to a nanoparticle 有权
标题翻译：用于定量与纳米颗粒缀合的生物分子的方法
公开(公告)号：US20080274463A1
公开(公告)日：2008-11-06
申请号：US11800360
申请日：2007-05-04
申请人： Xiao-Bo Chen , Christopher Bieniarz , Michael Farrell
发明人： Xiao-Bo Chen , Christopher Bieniarz , Michael Farrell
IPC分类号： C12Q1/68 , G01N1/00 , G01N33/00 , C12Q1/00 , G01N33/53 , G01N33/536 , G01N33/563 , G01N33/20 , C12Q1/37 , G01N21/76
CPC分类号： C12Q1/37 , B82Y15/00 , G01N33/54313 , G01N33/582 , Y10T436/143333 , Y10T436/17 , Y10T436/25125
摘要： Disclosed embodiments concern quantifying a biomolecule conjugated to a nanoparticle. Quantifying typically comprises determining the number of biomolecules per nanoparticle. Any suitable biomolecule can be used, including but not limited to, amino acids, peptides, proteins, haptens, nucleic acids, oligonucleotides, DNA, RNA, and combinations thereof. A single type of biomolecule may be conjugated to the nanoparticle, more than one biomolecule of a particular class may be conjugated to the nanoparticle, or two or more classes of biomolecules may be conjugated to the nanoparticle. Certain disclosed embodiments comprise enzymatically or chemically digesting a biomolecule conjugated to the nanoparticle, or displacing a biomolecule using ligand-exchange chemistry. Where biomolecule concentrations are determined, any technique suitable for determining biomolecule concentration can be used, such as spectrophotometric techniques, including measuring tryptophan fluorescence and using a standard fluorescence intensity versus biomolecule concentration curve.
摘要翻译：公开的实施方案涉及量化与纳米颗粒缀合的生物分子。量化通常包括确定每个纳米颗粒的生物分子的数量。可以使用任何合适的生物分子，包括但不限于氨基酸，肽，蛋白质，半抗原，核酸，寡核苷酸，DNA，RNA及其组合。单一类型的生物分子可以与纳米颗粒结合，特定类别的多于一个的生物分子可以与纳米颗粒缀合，或者两种或更多类别的生物分子可以与纳米颗粒缀合。某些公开的实施方案包括酶或化学消化与纳米颗粒缀合的生物分子，或使用配体交换化学位移生物分子。在确定生物分子浓度的情况下，可以使用适合于测定生物分子浓度的任何技术，例如分光光度技术，包括测量色氨酸荧光并使用标准荧光强度与生物分子浓度曲线。

3. 发明授权

US07682789B2 Method for quantifying biomolecules conjugated to a nanoparticle 有权
标题翻译：用于定量与纳米颗粒缀合的生物分子的方法
公开(公告)号：US07682789B2
公开(公告)日：2010-03-23
申请号：US11800360
申请日：2007-05-04
申请人： Xiao-Bo Chen , Christopher Bieniarz , Michael Farrell
发明人： Xiao-Bo Chen , Christopher Bieniarz , Michael Farrell
IPC分类号： C12Q1/68 , G01N33/53 , C07K14/00 , C07H21/00 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/37 , B82Y15/00 , G01N33/54313 , G01N33/582 , Y10T436/143333 , Y10T436/17 , Y10T436/25125
摘要： Disclosed embodiments concern quantifying a biomolecule conjugated to a nanoparticle. Quantifying typically comprises determining the number of biomolecules per nanoparticle. Any suitable biomolecule can be used, including but not limited to, amino acids, peptides, proteins, haptens, nucleic acids, oligonucleotides, DNA, RNA, and combinations thereof. A single type of biomolecule may be conjugated to the nanoparticle, more than one biomolecule of a particular class may be conjugated to the nanoparticle, or two or more classes of biomolecules may be conjugated to the nanoparticle. Certain disclosed embodiments comprise enzymatically or chemically digesting a biomolecule conjugated to the nanoparticle, or displacing a biomolecule using ligand-exchange chemistry. Where biomolecule concentrations are determined, any technique suitable for determining biomolecule concentration can be used, such as spectrophotometric techniques, including measuring tryptophan fluorescence and using a standard fluorescence intensity versus biomolecule concentration curve.
摘要翻译：公开的实施方案涉及量化与纳米颗粒缀合的生物分子。量化通常包括确定每个纳米颗粒的生物分子的数量。可以使用任何合适的生物分子，包括但不限于氨基酸，肽，蛋白质，半抗原，核酸，寡核苷酸，DNA，RNA及其组合。单一类型的生物分子可以与纳米颗粒结合，特定类别的多于一种生物分子可以与纳米颗粒缀合，或者两种或更多类别的生物分子可以与纳米颗粒缀合。某些公开的实施方案包括酶或化学消化与纳米颗粒缀合的生物分子，或使用配体交换化学位移生物分子。在确定生物分子浓度的情况下，可以使用适合于测定生物分子浓度的任何技术，例如分光光度技术，包括测量色氨酸荧光并使用标准荧光强度与生物分子浓度曲线。

4. 发明申请

US20100297725A1 Haptens, hapten conjugates, compositions thereof and method for their preparation and use 有权
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
IPC分类号： C12N9/96 , C07K16/00
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.

5. 发明授权

US09040310B2 Antibody-nanoparticle conjugates and methods for making and using such conjugates 有权
标题翻译：抗体 - 纳米颗粒缀合物和制备和使用这种缀合物的方法
公开(公告)号：US09040310B2
公开(公告)日：2015-05-26
申请号：US13640944
申请日：2011-04-27
申请人： Julia Ashworth-Sharpe , Chol Steven Yun , Zhanna Zhilina , Adrian E. Murillo , Donald D. Johnson , Michael Farrell , Jerome W. Kosmeder , Christopher Bieniarz
发明人： Julia Ashworth-Sharpe , Chol Steven Yun , Zhanna Zhilina , Adrian E. Murillo , Donald D. Johnson , Michael Farrell , Jerome W. Kosmeder , Christopher Bieniarz
IPC分类号： G01N33/544 , G01N33/68 , C07F9/12 , C07F9/50 , C07F9/572 , C07F9/655 , G01N33/531 , G01N33/543 , G01N33/58
CPC分类号： G01N33/54346 , C07F9/12 , C07F9/5004 , C07F9/5022 , C07F9/5045 , C07F9/5728 , C07F9/65515 , C07F9/65517 , C07F9/65522 , G01N33/531 , G01N33/553 , G01N33/587 , G01N33/6854 , G01N2333/916
摘要： Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein.
摘要翻译：本文公开了抗体 - 纳米颗粒缀合物，其包括直接连接到抗体或其片段的两个或更多个纳米颗粒（例如金，钯，铂，银，铜，镍，钴，铱或其两种或多种的合金）金属硫醇键。制备本文公开的抗体 - 纳米颗粒缀合物的方法包括使芳基膦 - 纳米颗粒复合物与还原抗体反应以产生抗体 - 纳米颗粒缀合物。本文还公开了用于检测样品中靶分子的方法，其包括使用抗体 - 纳米颗粒缀合物（例如本文所述的抗体 - 纳米颗粒缀合物）和利用本文公开的方法检测靶分子的试剂盒。

6. 发明授权

US08703417B2 Enzyme-catalyzed metal deposition for the enhanced detection of analytes of interest 有权
标题翻译：酶催化的金属沉积用于增强感兴趣的分析物的检测
公开(公告)号：US08703417B2
公开(公告)日：2014-04-22
申请号：US13530001
申请日：2012-06-21
申请人： Christopher Bieniarz , Michael Farrell
发明人： Christopher Bieniarz , Michael Farrell
IPC分类号： C12Q1/68 , G01N33/53
CPC分类号： C07F9/093 , C07F9/65517 , C07F9/65522 , C07H15/20 , C07H15/24 , C07H15/26 , G01N33/581
摘要： The invention is directed to enhanced methods for detecting an analyte of interest in situ, by immunoassay, or by hybridization comprising binding an enzyme-labeled conjugate molecule to an analyte of interest in the presence of a redox-inactive reductive species and a soluble metal ion. The enzyme catalyzes the conversion of the inactive reductive species to an active reducing agent, which in turn reduces the metal ion to a metal atom thereby providing an enhanced means of detecting the analyte via metal deposition.
摘要翻译：本发明涉及通过免疫测定或通过杂交来检测目的分析物的增强方法，包括在氧化还原惰性还原物质和可溶性金属离子存在下将酶标记的缀合物分子与目的分析物结合。酶催化无活性还原物质转化为活性还原剂，其又将金属离子还原成金属原子，从而提供通过金属沉积检测分析物的增强方法。

7. 发明申请

US20100105566A1 Enzyme-Catalyzed Metal Deposition for the Enhanced Detection of Analytes of Interest 有权
标题翻译：酶催化金属沉积用于增强感兴趣的分析物的检测
公开(公告)号：US20100105566A1
公开(公告)日：2010-04-29
申请号：US12608261
申请日：2009-10-29
申请人： Christopher Bieniarz , Michael Farrell
发明人： Christopher Bieniarz , Michael Farrell
IPC分类号： C40B30/00 , G01N33/53
CPC分类号： C07F9/093 , C07F9/65517 , C07F9/65522 , C07H15/20 , C07H15/24 , C07H15/26 , G01N33/581
摘要： The invention is directed to enhanced methods for detecting an analyte of interest in situ, by immunoassay, or by hybridization comprising binding an enzyme-labeled conjugate molecule to an analyte of interest in the presence of a redox-inactive reductive species and a soluble metal ion. The enzyme catalyzes the conversion of the inactive reductive species to an active reducing agent, which in turn reduces the metal ion to a metal atom thereby providing an enhanced means of detecting the analyte via metal deposition.
摘要翻译：本发明涉及通过免疫测定或通过杂交来检测目的分析物的增强方法，包括在氧化还原惰性还原物质和可溶性金属离子存在下将酶标记的缀合物分子与目的分析物结合。酶催化无活性还原物质转化为活性还原剂，其又将金属离子还原成金属原子，从而提供通过金属沉积检测分析物的增强方法。

8. 发明申请

US20080268462A1 Haptens, hapten conjugates, compositions thereof and method for their preparation and use 有权
标题翻译：半抗原，半抗原共轭物，其组合物及其制备和使用方法
公开(公告)号：US20080268462A1
公开(公告)日：2008-10-30
申请号：US11982627
申请日：2007-11-01
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
IPC分类号： G01N33/53 , C07K16/44 , C07C205/06 , C07D493/04 , C07H21/04 , C12N9/08 , C07D307/78
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.
摘要翻译：描述了用于执行多路诊断测定的方法，例如样品中两种或更多种不同靶标。一个实施方案包括使样品与特异性结合两个或更多个不同靶标的两个或多个特异性结合部分接触。两个或更多个特异性结合部分与不同的半抗原结合，并且至少一个半抗原是恶唑，吡唑，噻唑，除二硝基苯基以外的硝基芳基化合物，苯并呋咱，三萜，脲，硫脲，旋叶，香豆素，环链烷醇，杂二芳基，偶氮芳基或苯并二氮杂。将样品与两种或更多种可分别检测的不同的抗半抗原抗体接触。两种或更多种不同的抗半抗原抗体可以缀合到不同的可检测标记上。

9. 发明授权

US08481270B2 Method for chromogenic detection of two or more target molecules in a single sample 有权
标题翻译：一种样品中两种或更多种靶分子的显色检测方法
公开(公告)号：US08481270B2
公开(公告)日：2013-07-09
申请号：US13059274
申请日：2009-08-21
申请人： Richard Gniewek , Michael Farrell , Hiroaki Nitta , Megan Lehrkamp , Jerome Kosmeder , Brian Daniel Kelly , Thomas Grogan , Fabien Gaire , Mary Padilla , Christopher Bieniarz
发明人： Richard Gniewek , Michael Farrell , Hiroaki Nitta , Megan Lehrkamp , Jerome Kosmeder , Brian Daniel Kelly , Thomas Grogan , Fabien Gaire , Mary Padilla , Christopher Bieniarz
IPC分类号： C12Q1/68 , G01N33/58 , G01N33/53
CPC分类号： C12Q1/6841 , G01N33/53 , G01N33/581 , G01N33/583
摘要： The present invention provides a method and kit for detection of two or more target molecules in a single tissue sample, such as for gene and protein dual detection in a single tissue sample. Methods comprise treating a tissue sample with a first binding moiety that specifically binds a first target molecule. Methods further comprise treating the tissue sample with a solution containing a soluble electron-rich aromatic compound prior to or concomitantly with contacting the tissue sample with a hapten-labeled binding moiety and detecting a second target molecule. In one example, the first target molecule is a protein and the second is a nucleic acid sequence, the first target molecule being detected by immunohistochemistry and the second by in situ hybridization. The disclosed method reduces background due to non-specific binding of the hapten-labeled specific binding moiety to an insoluble electron rich compound deposited near the first target molecule.
摘要翻译：本发明提供了用于检测单个组织样品中两种或更多种靶分子的方法和试剂盒，例如用于单一组织样品中的基因和蛋白质双重检测。方法包括用特异性结合第一靶分子的第一结合部分处理组织样品。方法还包括在使组织样品与半抗原标记的结合部分接触或同时检测第二靶分子之前或同时使用含有可溶于富电子的芳族化合物的溶液处理组织样品。在一个实例中，第一靶分子是蛋白质，第二个是核酸序列，第一个靶分子通过免疫组织化学检测，第二个是原位杂交。所公开的方法由于半抗原标记的特异性结合部分与沉积在第一靶分子附近的不溶性富电子化合物的非特异性结合而减少背景。

10. 发明申请

US20100184087A1 Haptens, hapten conjugates, compositions thereof and method for their preparation and use 有权
标题翻译：半抗原，半抗原共轭物，其组合物及其制备和使用方法
公开(公告)号：US20100184087A1
公开(公告)日：2010-07-22
申请号：US12660017
申请日：2010-02-17
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
IPC分类号： G01N33/53
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.
摘要翻译：描述了用于执行多路诊断测定的方法，例如样品中两种或更多种不同靶标。一个实施方案包括使样品与特异性结合两个或更多个不同靶标的两个或多个特异性结合部分接触。两个或更多个特异性结合部分与不同的半抗原结合，至少一个半抗原是恶唑，吡唑，噻唑，除二硝基苯基以外的硝基芳基化合物，苯并呋咱，三萜烯，脲，硫脲，旋叶，香豆素，环链烷醇，杂二芳基，偶氮芳基或苯并二氮杂。将样品与两种或更多种可分别检测的不同的抗半抗原抗体接触。两种或更多种不同的抗半抗原抗体可以缀合到不同的可检测标记上。

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