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    Compositions and methods for immunoaffinity purification
    72.
    发明授权
    Compositions and methods for immunoaffinity purification 有权
    免疫亲和纯化的组合物和方法

    公开(公告)号：US07534861B2

    公开(公告)日：2009-05-19

    申请号：US10340179

    申请日：2003-01-10

    申请人： Kenneth J. Rothschild ,  Sadanand Gite ,  Jerzy Olejnik ,  Mark Lim

    发明人： Kenneth J. Rothschild ,  Sadanand Gite ,  Jerzy Olejnik ,  Mark Lim

    IPC分类号： C07K14/00 ,  C12N15/00

    CPC分类号： C07K14/4746 ,  C07K2319/20 ,  C07K2319/50 ,  C07K2319/70

    摘要： A hybrid polypeptide composed of a p53 epitope peptide and a desired functional protein are produced by recombinant DNA techniques. A DNA expression vector is constructed that includes segments of DNA coding for the epitope peptide and the desired functional protein. An optional linking portion is contemplated. The linking portion of the epitope peptide is cleavable at a specific amino acid residue adjacent the functional protein by use of a sequence specific proteolytic enzyme or chemical proteolytic agent. The hybrid polypeptide expressed by the host cells transformed by the cloning vector is removed therefrom and purified by affinity chromatography techniques by use of an immobilized antibody specific to the antigenic portion of the epitope peptide. The protein is then cleaved from the isolated hybrid polypeptide with an appropriate proteolic enzyme or chemical agent, thereby releasing the mature functional protein in highly purified, highly active state.

    摘要翻译： 通过重组DNA技术产生由p53表位肽和所需功能性蛋白质组成的杂合多肽。 构建DNA表达载体，其包括编码表位肽和所需功能性蛋白质的DNA片段。 预期可选的连接部分。 表位肽的连接部分可以通过使用序列特异性蛋白水解酶或化学蛋白水解剂在与功能蛋白相邻的特定氨基酸残基处可切割。 由克隆载体转化的宿主细胞表达的杂合多肽从其中除去，并通过使用对表位肽的抗原部分特异性的固定化抗体通过亲和层析技术进行纯化。 然后用合适的蛋白酶或化学试剂从分离的杂交多肽切割蛋白质，从而释放高度纯化，高活性状态的成熟功能性蛋白质。

    BIOREACTIVE AGENTS
    73.
    发明申请
    BIOREACTIVE AGENTS 审中-公开
    生物制剂

    公开(公告)号：US20090075253A1

    公开(公告)日：2009-03-19

    申请号：US11640800

    申请日：2006-12-18

    申请人： Kenneth J. Rothschild ,  Sanjay M. Sonar ,  Jerzy Olejnik

    发明人： Kenneth J. Rothschild ,  Sanjay M. Sonar ,  Jerzy Olejnik

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6876 ,  A61K38/00 ,  A61K41/0042 ,  C07C205/45 ,  C07C205/57 ,  C07D207/404 ,  C07D311/16 ,  C07D405/12 ,  C07D495/04 ,  C07H19/04 ,  C07H21/00 ,  C07K1/1077 ,  C07K1/13 ,  C12P21/00 ,  C12Q1/6823 ,  G01N33/5306 ,  G01N33/532 ,  G01N33/58 ,  C12Q2523/319

    摘要： This invention relates to agents and conjugates that can be used to detect and isolate target components from complex mixtures such as nucleic acids from biological samples, cells from bodily fluids, and nascent proteins from translation reactions. Agents comprise a detectable moiety bound to a photoreactive moiety. Conjugates comprise agents coupled to substrates by covalent bounds which can be selectively cleaved with the administration of electromagnetic radiation. Targets substances labeled with detectable molecules can be easily identified and separated from a heterologous mixture of substances. Exposure of the conjugate to radiation releases the target in a functional form and completely unaltered. Using photocleavable molecular precursors as the conjugates, label can be incorporated into macromolecules, the nascent macromolecules isolated and the label completely removed. The invention also relates to targets isolated with these conjugates which may be useful as pharmaceutical agents or compositions that can be administered to humans and other mammals. Useful compositions include biological agents such as nucleic acids, proteins, lipids and cytokines. Conjugates can also be used to monitor the pathway and half-life of pharmaceutical composition in vivo and for diagnostic, therapeutic and prophylactic purposes. The invention also relates to kits comprised of agents and conjugates that can be used for the detection of diseases, disorders and nearly any individual substance in a complex background of substances.

    摘要翻译： 本发明涉及可用于从复杂混合物检测和分离靶组分的试剂和缀合物，例如来自生物样品的核酸，来自体液的细胞和来自翻译反应的新生蛋白。 试剂包含与光反应性部分结合的可检测部分。 缀合物包括通过共价界限偶联到底物的试剂，其可以通过施用电磁辐射被选择性地切割。 目标可以用可检测分子标记的物质可以很容易地识别，并与物质的异源混合物分离。 缀合物暴露于放射线以功能形式释放靶，完全不变。 使用可光致发光的分子前体作为共轭物，标记可以并入大分子中，分离出新生大分子，并将标记完全除去。 本发明还涉及用这些缀合物分离的靶，其可用作可以施用于人和其它哺乳动物的药剂或组合物。 有用的组合物包括生物制剂如核酸，蛋白质，脂质和细胞因子。 缀合物也可用于监测药物组合物在体内和诊断，治疗和预防目的中的途径和半衰期。 本发明还涉及可用于检测疾病，病症和物质复杂背景中的几乎任何单一物质的试剂和缀合物的试剂盒。

    Detection of markers in nascent proteins
    74.
    发明授权
    Detection of markers in nascent proteins 失效
    新生蛋白中标记物的检测

    公开(公告)号：US07211394B2

    公开(公告)日：2007-05-01

    申请号：US11145781

    申请日：2005-06-06

    申请人： Kenneth J. Rothschild ,  Jerzy Olejnik ,  Sanjay M. Sonar

    发明人： Kenneth J. Rothschild ,  Jerzy Olejnik ,  Sanjay M. Sonar

    IPC分类号： C12Q1/68

    CPC分类号： C07D207/404 ,  A61K38/00 ,  A61K41/0042 ,  C07C205/45 ,  C07C205/57 ,  C07D311/16 ,  C07D405/12 ,  C07D495/04 ,  C07H19/04 ,  C07H21/00 ,  C07K1/1077 ,  C07K1/13 ,  C12P21/00 ,  C12Q1/68 ,  G01N33/532 ,  G01N33/582

    摘要： The invention is directed to methods for the non-radioactive labeling, detection, quantitation and isolation of nascent proteins translated in a cellular or cell-free translation system. tRNA molecules are misaminoacylated with non-radioactive markers which may be non-native amino acids, amino acid analogs or derivatives, or substances recognized by the protein synthesizing machinery. Markers may comprise cleavable moieties, detectable labels, reporter properties wherein markers incorporated into protein can be distinguished from unincorporated markers, or coupling agents which facilitate the detection and isolation of nascent protein from other components of the translation system. The invention also comprises proteins prepared using misaminoacylated tRNAs which can be utilized in pharmaceutical compositions for the treatment of diseases and disorders in humans and other mammals, and kits which may be used for the detection of diseases and disorders.

    摘要翻译： 本发明涉及用于在细胞或无细胞翻译系统中翻译的新生蛋白的非放射性标记，检测，定量和分离的方法。 tRNA分子被非放射性标记物错氨基酰化，其可以是非天然氨基酸，氨基酸类似物或衍生物，或由蛋白质合成机理识别的物质。 标记可以包含可切割部分，可检测标记，报告基因特性，其中掺入蛋白质的标记可以与非组合标记物或促进从翻译系统的其他组分检测和分离新生蛋白质的偶联剂区别。 本发明还包括使用可用于治疗人类和其他哺乳动物疾病和病症的药物组合物中的使用错氨基酰化tRNA制备的蛋白质，以及可用于检测疾病和病症的试剂盒。

    Detection of disease related genes
    75.
    发明授权
    Detection of disease related genes 有权
    检测疾病相关基因

    公开(公告)号：US07169558B2

    公开(公告)日：2007-01-30

    申请号：US10401251

    申请日：2003-03-27

    申请人： Kenneth J. Rothschild ,  Sanjay M. Sonar ,  Jerzy Olejnik

    发明人： Kenneth J. Rothschild ,  Sanjay M. Sonar ,  Jerzy Olejnik

    IPC分类号： C12Q1/68

    CPC分类号： A61K41/0042 ,  A61K38/00 ,  C07C205/45 ,  C07C205/57 ,  C07D207/404 ,  C07D311/16 ,  C07D405/12 ,  C07D495/04 ,  C07H19/04 ,  C07H21/00 ,  C07K1/1077 ,  C07K1/13 ,  C12P21/00 ,  C12Q1/6883 ,  G01N33/58

    摘要： The invention is directed to methods for the non-radioactive labeling, detection, quantitation and isolation of nascent proteins translated in a cellular or cell-free translation system. tRNA molecules are misaminoacylated with non-radioactive markers which may be non-native amino acids, amino acid analogs or derivatives, or substances recognized by the protein synthesizing machinery. Markers may comprise cleavable moieties, detectable labels, reporter properties wherein markers incorporated into protein can be distinguished from unincorporated markers, or coupling agents which facilitate the detection and isolation of nascent protein from other components of the translation system. The invention also comprises proteins prepared using misaminoacylated tRNAs which can be utflized in pharmaceutical compositions for the treatment of diseases and disorders in humans and other maninials, and kits which may be used for the detection of diseases and disorders.

    摘要翻译： 本发明涉及用于在细胞或无细胞翻译系统中翻译的新生蛋白的非放射性标记，检测，定量和分离的方法。 tRNA分子被非放射性标记物错氨基酰化，其可以是非天然氨基酸，氨基酸类似物或衍生物，或由蛋白质合成机理识别的物质。 标记可以包含可切割部分，可检测标记，报告基因特性，其中掺入蛋白质的标记可以与非组合标记物或促进从翻译系统的其他组分检测和分离新生蛋白质的偶联剂区别。 本发明还包括使用错氨基酰化tRNA制备的蛋白质，其可以在药物组合物中被用于治疗人类和其他肌肉疾病和病症，以及可用于检测疾病和病症的试剂盒。

    Methods for the detection, analysis and isolation of Nascent proteins
    79.
    发明授权
    Methods for the detection, analysis and isolation of Nascent proteins 有权
    检测，分析和分离Nascent蛋白的方法

    公开(公告)号：US06306628B1

    公开(公告)日：2001-10-23

    申请号：US09382736

    申请日：1999-08-25

    申请人： Kenneth J. Rothschild ,  Sadanand Gite ,  Jerzy Olejnik

    发明人： Kenneth J. Rothschild ,  Sadanand Gite ,  Jerzy Olejnik

    IPC分类号： C12P1934

    CPC分类号： C12P21/00 ,  C12N15/10 ,  C12N15/67 ,  C12P21/02 ,  G01N33/6803

    摘要： This invention relates to non-radioactive markers that facilitate the detection and analysis of nascent proteins translated within cellular or cell-free translation systems. Nascent proteins containing these markers can be rapidly and efficiently detected, isolated and analyzed without the handling and disposal problems associated with radioactive reagents. Preferred markers are dipyrrometheneboron difluoride (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) dyes.

    摘要翻译： 本发明涉及促进在细胞或无细胞翻译系统中翻译的新生蛋白的检测和分析的非放射性标记物。 可以快速有效地检测，分离和分析含有这些标记物的蛋白质，而不会与放射性试剂相关的处理和处置问题。 优选的标记是二吡咯亚甲基二氟化物（4,4-二氟-4-硼酸-3a，4a-二氮杂 - 顺 - 并苯并三嗪）染料。