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。

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。

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。

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。

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。

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。

公开(公告)号：US20060141554A1

公开(公告)日：2006-06-29

申请号：US10966536

申请日：2004-10-14

申请人： Kyle Gee ,  Courtenay Hart ,  Richard Haugland ,  Wayne Patton ,  Scott Whitney

发明人： Kyle Gee ,  Courtenay Hart ,  Richard Haugland ,  Wayne Patton ,  Scott Whitney

IPC分类号： G01N33/48 ,  C07F15/04

CPC分类号： C07D263/62 ,  A61K49/0021 ,  A61K49/0039 ,  A61K49/0041 ,  A61K49/0052 ,  C07D311/12 ,  C07D311/16 ,  C07D311/18 ,  C07D405/14 ,  C07F5/022 ,  G01N1/30 ,  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.

公开(公告)号：US20060160068A1

公开(公告)日：2006-07-20

申请号：US10943463

申请日：2004-09-17

申请人： Joseph Beechem ,  Kyle Gee ,  David Hagen ,  Iain Johnson ,  Hee-Chol Kang ,  Christina Pastula

发明人： Joseph Beechem ,  Kyle Gee ,  David Hagen ,  Iain Johnson ,  Hee-Chol Kang ,  Christina Pastula

IPC分类号： C12Q1/70 ,  C12Q1/68 ,  C07K16/46 ,  C07F5/02 ,  C07D311/88

CPC分类号： G01N33/542 ,  C07F9/12 ,  C07F9/65318 ,  C07F9/65522 ,  C07F9/6596 ,  C07K16/46 ,  Y10S435/968 ,  Y10S436/80 ,  Y10S530/807 ,  Y10T436/13

摘要： The present invention provides ligand-detection reagents, ligand analogs and methods for determining the presence of a ligand in a sample. The ligand-detection reagent comprises a ligand-binding antibody and a ligand analog to form an antibody-ligand analog complex wherein the ligand analog is covalently bonded to a reporter molecule. This complex may additionally comprise a labeling protein non-covalently bonded to the antibody to form a ternary complex wherein the labeling protein comprises a monovalent antibody fragment or a non-antibody protein that is covalently bonded to a label moiety. The reporter molecule is either quenched by the ligand-binding antibody or by the label moiety of the labeling protein, depending on the reporter molecule and the ligand-binding antibody, wherein the amount of quenching is directly related to the amount of ligand present in the sample. Alternatively, the ligand analog is fluorogenic wherein the ligand analog is essentially non-fluorescent in solution but when bound by the ligand-binding antibody the detectable signal increases. In this instance a decrease in signal, as opposed to the relieving of quenching, is measured for the presence of a target ligand.

摘要翻译： 本发明提供配体检测试剂，配体类似物和用于测定样品中配体存在的方法。 配体检测试剂包含配体结合抗体和配体类似物以形成抗体 - 配体类似物复合物，其中配体类似物共价键合到报告分子。 该复合物还可以包含与抗体非共价键合以形成三元复合物的标记蛋白质，其中标记蛋白质包含共价键合到标记部分的单价抗体片段或非抗体蛋白质。 取决于报道分子和配体结合抗体，报告分子被配体结合抗体或标记蛋白的标记部分淬灭，其中淬灭量与存在于 样品。 或者，配体类似物是荧光的，其中配体类似物在溶液中基本上是非荧光的，但当被配体结合抗体结合时，可检测信号增加。 在这种情况下，与目标配体的存在相比，测量了与消除淬灭相反的信号降低。

公开(公告)号：US20060063269A1

公开(公告)日：2006-03-23

申请号：US11157467

申请日：2005-06-20

申请人： Brian Agnew ,  Kyle Gee

发明人： Brian Agnew ,  Kyle Gee

IPC分类号： G01N33/48

CPC分类号： G01N33/582 ,  C09B11/24 ,  G01N33/533 ,  G01N33/534 ,  G01N33/583 ,  G01N33/60 ,  G01N33/6848

摘要： The present invention provides novel reactive fluorescent compounds that incorporate stable isotopic (deuterium, 13-carbon, 15-nitrogen, 18-oxygen) substitutions. The invention includes the use of these compounds, in combination with non-isotopically substituted analogs, for the purification, identification and relative quantification of proteins, peptides, saccharides, metabolites, and other biologically important compounds by combining liquid chromatography (LC) and mass spectrometry (MS). Fluorescent labeling of target compounds in this manner provides orders-of-magnitude sensitivity enhancement over traditional stable isotope labels, and also affords the possibility of simultaneous multiplexed analysis due to the multiwavelength nature of different fluorophores.

摘要翻译： 本发明提供了引入稳定同位素（氘，13-碳，15-氮，18-氧）取代的新型反应性荧光化合物。 本发明包括将这些化合物与非同位素取代的类似物组合用于通过液相色谱（LC）和质谱法的组合来纯化，鉴定和相对定量蛋白质，肽，糖，代谢物和其它生物学上重要的化合物 （女士）。 目标化合物的荧光标记以这种方式提供超过传统稳定同位素标记的数量级的灵敏度增强，并且由于不同荧光团的多波长性质，也提供同时多重分析的可能性。

公开(公告)号：US20050014197A1

公开(公告)日：2005-01-20

申请号：US10821522

申请日：2004-04-09

申请人： Brian Agnew ,  Kyle Gee ,  Vladimir Martin

发明人： Brian Agnew ,  Kyle Gee ,  Vladimir Martin

IPC分类号： C12Q1/48 ,  G01N33/53

CPC分类号： 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。