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    • 7. 发明申请
    • METHODS AND COMPOSITIONS RELATED TO MODULATORS OF EUKARYOTIC CELLS
    • 与真核细胞调节剂相关的方法和组合物
    • WO2014035693A2
    • 2014-03-06
    • PCT/US2013/055362
    • 2013-08-16
    • THE SCRIPPS RESEARCH INSTITUTEZHANG, HongkaiWILSON, Ian, A.LERNER, Richard, A.
    • ZHANG, HongkaiWILSON, Ian, A.LERNER, Richard, A.
    • C40B30/06
    • C07K16/2863C07K14/71C07K14/7153C07K16/2839C07K16/2866C07K2317/21C07K2317/31C07K2317/35C07K2317/622C07K2317/74C07K2317/75C07K2317/92C07K2319/30C40B30/06G01N33/6845G01N33/6854G01N33/746
    • The invention provides methods for identifying protein modulators (e.g., antibody agonists) of eukaryotic cells. The methods typically involve expressing a combinatorial agent library (e.g., via lentiviral vectors) inside a eukaryotic cell type (e.g., a mammalian cell) and then directly selecting for agents (e.g., antibodies) that are agonist, of a target molecule (e.g.,. a signaling receptor) that modulates a phenotype of or elicits a cellular response in the cell. Some related methods involve co-culturing a cell expressing a combinatorial agent library and a second cell, and then selecting agents that modulate a phenotype of or elicit a cellular response in the second cell. Preferably, the agents are antibodies and are introduced into and expressed in the starting cells under conditions each individual cell expresses no more than 3 different members of the antibody library. In addition, the invention provides methods for identifying protein agonists that capable of reprograming or trans-differentiating a target cell. Also provided in the invention are specific agonist antibodies of signaling receptors or biomolecules that modulate a phenotype or effectuate a cellular response in a eukaryotic cell (e.g., agonist antibodies of EpoR, TpoR or G-CSFR). Further provided in the invention are methods for selecting from combinatorial antibody libraries bispecific antibodies that can regulate cell phenotypes.
    • 本发明提供鉴定真核细胞的蛋白质调节剂(例如抗体激动剂)的方法。 方法通常涉及在真核细胞类型(例如,哺乳动物细胞)内表达组合试剂文库(例如通过慢病毒载体),然后直接选择靶分子的激动剂(例如,抗体)(例如, 一种信号受体),其调节细胞中的表型或引起细胞反应。 一些相关方法涉及共培养表达组合试剂文库和第二细胞的细胞,然后选择调节第二细胞中表型或引起细胞应答的试剂。 优选地,所述试剂是抗体,并且在每个单个细胞表达不超过3个抗体文库的不同成员的条件下被引入并在起始细胞中表达。 此外,本发明提供了鉴定能够重新编程或反向分化靶细胞的蛋白质激动剂的方法。 本发明还提供了调节表型或在真核细胞(例如EpoR,TpoR或G-CSFR的激动剂抗体)中调节细胞应答的信号传导受体或生物分子的特异性激动剂抗体。 本发明还提供了从组合抗体文库中选择可以调节细胞表型的双特异性抗体的方法。
    • 8. 发明申请
    • BIOLUMINESCENT PLANTS AND METHODS OF MAKING SAME
    • 生物发光植物及其制备方法
    • WO2002081647A2
    • 2002-10-17
    • PCT/US2002/011116
    • 2002-04-08
    • THE SCRIPPS RESEARCH INSTITUTEKAY, Steve, A.KUHLMANN, TinaLERNER, Richard, A.
    • KAY, Steve, A.KUHLMANN, TinaLERNER, Richard, A.
    • C12N
    • C12N15/8241
    • A genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide, which is expressed in amount sufficient to produce at least about 750,000 photons of visible light/mm 2 /second, is provided, as is a visibly bioluminescent transgenic plant, which contains such a genetically modified plant cell. Also provided is a recombinant nucleic acid molecule, which contains a plant translational enhancer operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide. In addition, methods of producing a genetically modified plant cell that is visibly bioluminescent introducing a transgene encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm 2 /second are provided, as are kits containing such visibly bioluminescent compositions.
    • 提供了含有编码生物发光多肽的异源核苷酸序列的遗传修饰的植物细胞,其以足以产生至少约750000个可见光/ mm 2 /秒的光子的量表达,如可见生物发光转基因植物, 其含有这样的遗传修饰的植物细胞。 还提供了重组核酸分子,其包含与编码生物发光多肽的核苷酸序列可操作地连接的植物翻译增强子。 另外,生产显性生物发光的转基因植物细胞的方法是将编码生物发光多肽的转基因导入植物细胞,由此生物发光多肽以至少约75万光束的可见光/ mm 2表达 > /秒,以及含有这种可视生物发光组合物的试剂盒。
    • 10. 发明申请
    • ESCAPE LIBRARIES OF TARGET POLYPEPTIDES
    • ESCAPE目标聚合物图书馆
    • WO2009009045A2
    • 2009-01-15
    • PCT/US2008/008367
    • 2008-07-08
    • THE SCRIPPS RESEARCH INSTITUTELERNER, Richard, A.BRENNER, SydneyDICKERSON, Tobin, J.JANDA, Kim, D.
    • LERNER, Richard, A.BRENNER, SydneyDICKERSON, Tobin, J.JANDA, Kim, D.
    • C40B30/06
    • G01N33/6845C12N15/1037G01N2500/02
    • The present invention provides methods for identifying escape variants of a target polypeptide. The methods typically involve generating a library of variant polypeptides of the target polypeptide on a replicable display platform (e.g., phage display), identifying variant polypeptides which maintain the ability to bind to a known binding partner of the target polypeptide, and examining the identified variant polypeptides to identify escape variants whose binding to the binding partner is not antagonized by a library of known compounds which disrupt binding between the target polypeptide and the binding partner. The methods can further entail screening a library of candidate antagonist agents to identify a cognate antagonist agent which antagonizes binding between the escape variant and the binding partner. Additional escape variants can be identified by creating a library of further variant polypeptides of the identified escape variant, and subject the further variant polypeptides to the screening process. In each subsequent round of screening, the identified antagonist agent is combined with the library of known compounds employed in the previous round of screening. The invention also provides related methods for identifying novel antagonist agents of a target polypeptide. Further provided are vectors and kits for displaying a functional multimeric target protein on the surface of a phage.
    • 本发明提供了鉴定靶多肽逃避变体的方法。 所述方法通常涉及在可复制的显示平台(例如,噬菌体展示)上产生靶多肽的变体多肽文库,鉴定维持结合目标多肽的已知结合配偶体的能力的变体多肽,并检查鉴定的变体 用于鉴定与结合配偶体结合的逃避变体的多肽不被破坏靶多肽和结合配偶体之间的结合的已知化合物文库拮抗。 所述方法可进一步包括筛选候选拮抗剂试剂盒,以鉴定拮抗逃生变体与结合配偶体之间结合的同源拮抗剂。 可以通过产生所鉴定的逃避变体的另外的变体多肽的文库来鉴定另外的逃避变体,并将进一步的变体多肽进行筛选过程。 在随后的每一轮筛选中,将所鉴定的拮抗剂与前一轮筛选中使用的已知化合物的文库组合。 本发明还提供了鉴定目标多肽的新型拮抗剂的相关方法。 还提供了用于在噬菌体表面上显示功能性多聚体靶蛋白的载体和试剂盒。