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    Site-specific labeling of affinity tags in fusion proteins
    1.
    发明申请
    Site-specific labeling of affinity tags in fusion proteins 审中-公开
    融合蛋白中亲和标签的位点特异性标记

    公开(公告)号：US20050233307A1

    公开(公告)日：2005-10-20

    申请号：US10661451

    申请日：2003-09-12

    申请人： Kyle Gee ,  Courtenay Hart ,  Wai-Yee Leung ,  Wayne Patton ,  Aleksey Rukavishnikov ,  Richard Haugland ,  Zhenjun Diwu

    发明人： Kyle Gee ,  Courtenay Hart ,  Wai-Yee Leung ,  Wayne Patton ,  Aleksey Rukavishnikov ,  Richard Haugland ,  Zhenjun Diwu

    IPC分类号： A61K49/00 ,  C07D311/12 ,  C07D311/18 ,  C07F5/02 ,  C12Q1/00 ,  G01N20060101 ,  G01N33/00 ,  G01N33/52 ,  G01N33/533

    CPC分类号： G01N33/52 ,  A61K49/0021 ,  A61K49/0039 ,  A61K49/0041 ,  A61K49/0052 ,  C07D311/12 ,  C07D311/16 ,  C07D311/18 ,  C07F5/022 ,  G01N33/533 ,  G01N33/582

    摘要： The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a poly-histidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6. The acetic acid groups interact directly with the positively charged histidine residues of the affinity tag to effectively label and detect a fusion protein containing such an affinity tag when present in an acidic or neutral environment.

    摘要翻译： 本发明提供了通过能够选择性地结合亲和标签来促进融合蛋白的检测和标记的方法和荧光化合物。 荧光化合物具有通式A（B）n，其中A是荧光团，B是作为带电化学部分的结合结构域，其蛋白质或片段，n是1-6的整数，条件是 蛋白质或其片段不是抗体或从抗体产生。 本发明提供用于检测和标记含有多组氨酸亲和标签的融合蛋白的特异性荧光化合物和方法。 这些化合物具有通式A（L）m（B）n，其中A是荧光团，L是连接体，B是乙酸结合域，m是1至4的整数，n是1至 乙酸基团与亲和标签的带正电荷的组氨酸残基直接相互作用，以有效地标记和检测当存在于酸性或中性环境中时含有这种亲和标签的融合蛋白。

    SITE-SPECIFIC LABELING OF AFFINITY TAGS IN FUSION PROTEINS
    2.
    发明申请
    SITE-SPECIFIC LABELING OF AFFINITY TAGS IN FUSION PROTEINS 审中-公开
    融合蛋白中的亲和标签的特定标签

    公开(公告)号：US20090081722A1

    公开(公告)日：2009-03-26

    申请号：US12117689

    申请日：2008-05-08

    申请人： Kyle Gee ,  Courtenay Hart ,  Wai-Yee Leung ,  Wayne Patton ,  Aleksey Rukavishnikov ,  Richard Haugland ,  Zhenjun Diwu

    发明人： Kyle Gee ,  Courtenay Hart ,  Wai-Yee Leung ,  Wayne Patton ,  Aleksey Rukavishnikov ,  Richard Haugland ,  Zhenjun Diwu

    IPC分类号： G01N1/30 ,  C07F5/02 ,  C07D311/06

    CPC分类号： G01N33/52 ,  A61K49/0021 ,  A61K49/0039 ,  A61K49/0041 ,  A61K49/0052 ,  C07D311/12 ,  C07D311/16 ,  C07D311/18 ,  C07F5/022 ,  G01N33/533 ,  G01N33/582

    摘要： The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a poly-histidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6. The acetic acid groups interact directly with the positively charged histidine residues of the affinity tag to effectively label and detect a fusion protein containing such an affinity tag when present in an acidic or neutral environment.

    摘要翻译： 本发明提供了通过能够选择性地结合亲和标签来促进融合蛋白的检测和标记的方法和荧光化合物。 荧光化合物具有通式A（B）n，其中A是荧光团，B是作为带电化学部分的结合结构域，其蛋白质或片段，n是1-6的整数，条件是 蛋白质或其片段不是抗体或从抗体产生。 本发明提供用于检测和标记含有多组氨酸亲和标签的融合蛋白的特异性荧光化合物和方法。 这些化合物具有通式A（L）m（B）n，其中A是荧光团，L是连接体，B是乙酸结合域，m是1至4的整数，n是1至 乙酸基团与亲和标签的带正电荷的组氨酸残基直接相互作用，以有效地标记和检测当存在于酸性或中性环境中时含有这种亲和标签的融合蛋白。

    Reagents for labeling biomolecules having aldehyde or ketone moieties
    5.
    发明申请
    Reagents for labeling biomolecules having aldehyde or ketone moieties 有权
    用于标记具有醛或酮部分的生物分子的试剂

    公开(公告)号：US20050250152A1

    公开(公告)日：2005-11-10

    申请号：US11182122

    申请日：2005-07-15

    申请人： Richard Haugland ,  Thomas Steinberg ,  Wayne Patton ,  Zhenjun Diwu

    发明人： Richard Haugland ,  Thomas Steinberg ,  Wayne Patton ,  Zhenjun Diwu

    IPC分类号： G01N33/532 ,  C07D239/91 ,  C07D239/94 ,  C07D249/08 ,  C07D263/60 ,  C07D265/22 ,  C07D277/66 ,  C07D401/04 ,  G01N33/58 ,  C12Q1/68 ,  C07H21/04 ,  C08B37/00

    CPC分类号： C07D265/22 ,  C07D239/91 ,  Y10T436/143333 ,  Y10T436/200833

    摘要： Novel fluorescent derivitization reagents are described that are suitable for coupling to biomolecules that contain aldehyde or ketone functional groups. In one embodiment is provided reagents that have the following formula: wherein Q is carbonyl, thiocarbonyl, or sulfonyl, and R5 is -L-Z; L is arylene, or a C1-6 perfluoroalkylene, or a single covalent bond; Z is a carbonyl hydrazide, hydrazide, sulfonyl hydrazide, or a thiocarbonyl hydrazide; R11-R14 are independently H, C1-6alkyl, C1-6 alkoxy, C1-6 perfluoroalkyl, C1-6 alklyamino, di(C2-21-alkyl)amino, amino, carboxy, cyano, halogen, hydroxy, nitro, phenyl, or sulfo; and R21-R24 are independently H, C1-6 alkyl, C1-6 alkoxy, C1-6 perfluoroalkyl, C1-6 alklyamino, di(C2-12-alkyl)amino, amino, carboxy, cyano, halogen, hydroxy, nitro, phenyl, sulfo, or -L-Z. The method of treating a sample with the derivativization reagents is described. The reagents are particularly useful for labeling glycoproteins or glycopeptides, nucleic acids, and lipopolysaccharides in electrophoresis gels.

    摘要翻译： 描述了适用于与含有醛或酮官能团的生物分子偶联的新型荧光衍生试剂。 在一个实施方案中，提供具有下式的试剂：其中Q是羰基，硫代羰基或磺酰基，R 5是-L-Z; L是亚芳基或C 1-6个全氟亚烷基，或单个共价键; Z是羰基酰肼，酰肼，磺酰肼或硫代羰基酰肼; R 11 -R 14独立地为H，C 1-6烷基，C 1-6烷氧基，C 全氟烷基，C 1-6烷基氨基，二（C 1 -C 21 - 亚烷基）氨基，氨基，羧基，氰基，卤素 ，羟基，硝基，苯基或磺基; R 21和R 24独立地是H，C 1-6烷基，C 1-6烷氧基， C 1-6烷基全氟烷基，C 1-6烷基氨基，二（C 2-12-12 - 烷基）氨基，氨基，羧基，氰基， 卤素，羟基，硝基，苯基，磺基，或-LZ。 描述了用衍生化试剂处理样品的方法。 这些试剂特别可用于在电泳凝胶中标记糖蛋白或糖肽，核酸和脂多糖。

    Compositions and methods for detection and isolation of phosphorylated molecules
    6.
    发明申请
    Compositions and methods for detection and isolation of phosphorylated molecules 审中-公开
    用于磷酸化分子检测和分离的组合物和方法

    公开(公告)号：US20100009381A1

    公开(公告)日：2010-01-14

    申请号：US12500854

    申请日：2009-07-10

    申请人： Brian Agnew ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Thomas Steinberg ,  Wayne Patton

    发明人： Brian Agnew ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Thomas Steinberg ,  Wayne Patton

    IPC分类号： G01N33/53

    CPC分类号： G01N33/5008 ,  C12Q1/42 ,  C12Q1/485 ,  G01N33/502 ,  G01N33/5091 ,  G01N2333/4709 ,  Y10T436/142222 ,  Y10T436/145555 ,  Y10T436/147777 ,  Y10T436/201666

    摘要： The present invention relates to phosphate-binding compounds that find use in binding, detecting and isolating phosphorylated target molecules including the subsequent identification of target molecules that interact with phosphorylated target molecules or molecules capable of being phosphorylated. A binding solution is provide that comprises a phosphate-binding compound, an acid and a metal ion wherein the metal ion simultaneously interacts with an exposed phosphate group on a target molecule and the metal chelating moiety of the phosphate-binding compound forming a bridge between the phosphate-binding compound and a phosphorylated target molecule resulting in a ternary complex. The binding solution of the present invention finds use in binding and detecting immobilized and solubilized phosphorylated target molecules, isolation of phosphorylated target molecules from a complex mixture and aiding in proteomic analysis wherein kinase and phosphatase substrates and enzymes can be identified.

    摘要翻译： 本发明涉及用于结合，检测和分离磷酸化靶分子的磷酸盐结合化合物，包括随后鉴定与磷酸化靶分子或能够被磷酸化的分子相互作用的靶分子。 提供的结合溶液包括磷酸盐结合化合物，酸和金属离子，其中金属离子同时与靶分子上暴露的磷酸基团相互作用，并且磷酸盐结合化合物的金属螯合部分形成在 磷酸化结合化合物和导致三元复合物的磷酸化靶分子。 本发明的结合溶液用于结合和检测固定和溶解的磷酸化靶分子，从复杂混合物中分离磷酸化靶分子，并辅助蛋白质组分析，其中可以鉴定激酶和磷酸酶底物和酶。

    Compositions and methods for detection and isolation of phosphorylated molecules
    7.
    发明申请
    Compositions and methods for detection and isolation of phosphorylated molecules 审中-公开
    用于磷酸化分子检测和分离的组合物和方法

    公开(公告)号：US20110021372A1

    公开(公告)日：2011-01-27

    申请号：US12821003

    申请日：2010-06-22

    申请人： Brian AGNEW ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Thomas Steinberg ,  Wayne Patton

    发明人： Brian AGNEW ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Thomas Steinberg ,  Wayne Patton

    IPC分类号： C40B30/04 ,  C12Q1/68 ,  G01N33/68 ,  G01N33/566

    CPC分类号： G01N33/5008 ,  C12Q1/42 ,  C12Q1/485 ,  G01N33/502 ,  G01N33/5091 ,  G01N2333/4709 ,  Y10T436/142222 ,  Y10T436/145555 ,  Y10T436/147777 ,  Y10T436/201666

    摘要： The present invention relates to phosphate-binding compounds that find use in binding, detecting and isolating phosphorylated target molecules including the subsequent identification of target molecules that interact with phosphorylated target molecules or molecules capable of being phosphorylated. A binding solution is provide that comprises a phosphate-binding compound, an acid and a metal ion wherein the metal ion simultaneously interacts with an exposed phosphate group on a target molecule and the metal chelating moiety of the phosphate-binding compound forming a bridge between the phosphate-binding compound and a phosphorylated target molecule resulting in a ternary complex. The binding solution of the present invention finds use in binding and detecting immobilized and solubilized phosphorylated target molecules, isolation of phosphorylated target molecules from a complex mixture and aiding in proteomic analysis wherein kinase and phosphatase substrates and enzymes can be identified.

    摘要翻译： 本发明涉及用于结合，检测和分离磷酸化靶分子的磷酸盐结合化合物，包括随后鉴定与磷酸化靶分子或能够被磷酸化的分子相互作用的靶分子。 提供的结合溶液包括磷酸盐结合化合物，酸和金属离子，其中金属离子同时与靶分子上暴露的磷酸基团相互作用，并且磷酸盐结合化合物的金属螯合部分形成在 磷酸化结合化合物和导致三元复合物的磷酸化靶分子。 本发明的结合溶液用于结合和检测固定和溶解的磷酸化靶分子，从复杂混合物中分离磷酸化靶分子，并辅助蛋白质组分析，其中可以鉴定激酶和磷酸酶底物和酶。

    COMPOSITIONS AND METHODS FOR DETECTION AND ISOLATION OF PHOSPHORYLATED MOLECULES
    8.
    发明申请
    COMPOSITIONS AND METHODS FOR DETECTION AND ISOLATION OF PHOSPHORYLATED MOLECULES 审中-公开
    用于检测和分离磷酸化分子的组合物和方法

    公开(公告)号：US20080044804A1

    公开(公告)日：2008-02-21

    申请号：US11697103

    申请日：2007-04-05

    申请人： Brian AGNEW ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Thomas Steinberg ,  Wayne Patton

    发明人： Brian AGNEW ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Thomas Steinberg ,  Wayne Patton

    IPC分类号： G01N33/00 ,  C12Q1/00 ,  G01N33/20

    CPC分类号： G01N33/5008 ,  C12Q1/42 ,  C12Q1/485 ,  G01N33/502 ,  G01N33/5091 ,  G01N2333/4709 ,  Y10T436/142222 ,  Y10T436/145555 ,  Y10T436/147777 ,  Y10T436/201666

    摘要： The present invention relates to phosphate-binding compounds that find use in binding, detecting and isolating phosphorylated target molecules including the subsequent identification of target molecules that interact with phosphorylated target molecules or molecules capable of being phosphorylated. A binding solution is provide that comprises a phosphate-binding compound, an acid and a metal ion wherein the metal ion simultaneously interacts with an exposed phosphate group on a target molecule and the metal chelating moiety of the phosphate-binding compound forming a bridge between the phosphate-binding compound and a phosphorylated target molecule resulting in a ternary complex. The binding solution of the present invention finds use in binding and detecting immobilized and solubilized phosphorylated target molecules, isolation of phosphorylated target molecules from a complex mixture and aiding in proteomic analysis wherein kinase and phosphatase substrates and enzymes can be identified.

    摘要翻译： 本发明涉及用于结合，检测和分离磷酸化靶分子的磷酸盐结合化合物，包括随后鉴定与磷酸化靶分子或能够被磷酸化的分子相互作用的靶分子。 提供的结合溶液包括磷酸盐结合化合物，酸和金属离子，其中金属离子同时与靶分子上暴露的磷酸基团相互作用，并且磷酸盐结合化合物的金属螯合部分形成在 磷酸化结合化合物和导致三元复合物的磷酸化靶分子。 本发明的结合溶液用于结合和检测固定和溶解的磷酸化靶分子，从复杂混合物中分离磷酸化靶分子，并辅助蛋白质组分析，其中可以鉴定激酶和磷酸酶底物和酶。

    Compositions and methods for detection and isolation of phosphorylated molecules
    9.
    发明授权
    Compositions and methods for detection and isolation of phosphorylated molecules 有权
    用于磷酸化分子检测和分离的组合物和方法

    公开(公告)号：US07102005B2

    公开(公告)日：2006-09-05

    申请号：US10428192

    申请日：2003-05-02

    申请人： Brian Agnew ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Jixiang Liu ,  Vladimir Martin ,  Wayne Patton ,  Thomas Steinberg

    发明人： Brian Agnew ,  Joseph Beechem ,  Kyle Gee ,  Richard Haugland ,  Jixiang Liu ,  Vladimir Martin ,  Wayne Patton ,  Thomas Steinberg

    IPC分类号： C07D239/88 ,  C07D277/62 ,  C07D311/82 ,  C07D417/02 ,  C07D495/04

    CPC分类号： G01N33/50 ,  G01N33/5008 ,  G01N33/502 ,  G01N33/5091 ,  G01N2333/4709

    摘要： The present invention relates to phosphate-binding compounds that find use in binding, detecting and isolating phosphorylated target molecules including the subsequent identification of target molecules that interact with phosphorylated target molecules or molecules capable of being phosphorylated. A binding solution is provide that comprises a phosphate-binding compound, an acid and a metal ion wherein the metal ion simultaneously interacts with an exposed phosphate group on a target molecule and the metal chelating moiety of the phosphate-binding compound forming a bridge between the phosphate-binding compound and a phosphorylated target molecule resulting in a ternary complex. The binding solution of the present invention finds use in binding and detecting immobilized and solubilized phosphorylated target molecules, isolation of phosphorylated target molecules from a complex mixture and aiding in proteomic analysis wherein kinase and phosphatase substrates and enzymes can be identified.

    摘要翻译： 本发明涉及用于结合，检测和分离磷酸化靶分子的磷酸盐结合化合物，包括随后鉴定与磷酸化靶分子或能够被磷酸化的分子相互作用的靶分子。 提供的结合溶液包括磷酸盐结合化合物，酸和金属离子，其中金属离子同时与靶分子上暴露的磷酸基团相互作用，并且磷酸盐结合化合物的金属螯合部分形成在 磷酸化结合化合物和导致三元复合物的磷酸化靶分子。 本发明的结合溶液用于结合和检测固定和溶解的磷酸化靶分子，从复杂混合物中分离磷酸化靶分子，并辅助蛋白质组分析，其中可以鉴定激酶和磷酸酶底物和酶。