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    Mammalian expression system
    2.
    发明授权
    Mammalian expression system 有权

    公开(公告)号：US11767537B2

    公开(公告)日：2023-09-26

    申请号：US15754881

    申请日：2016-08-25

    申请人： LONZA BIOLOGICS PLC.

    发明人： Robert Young ,  James D. Budge ,  Mark C. Smales

    IPC分类号： C12N15/85 ,  C07K14/005 ,  C07K16/00 ,  C12N5/071 ,  C12N7/00 ,  C12N9/12 ,  C12N15/67

    CPC分类号： C12N15/85 ,  C07K14/005 ,  C07K16/00 ,  C12N5/0682 ,  C12N7/00 ,  C12N9/1229 ,  C12N15/67 ,  C12Y207/04004 ,  C12N2800/108

    摘要： The present invention relates to an expression system for the heterologous expression of a nucleic acid sequence of interest in a mammalian cell, the system comprising: (i) a first genetic entity, comprising: a nucleic acid sequence encoding a functional Epstein Barr virus nuclear antigen 1 (EBNA-1), the nucleic acid sequence being operably linked to regulatory elements that allow for expression of the nucleic acid sequence encoding a functional EBNA-1; (ii) a second genetic entity, comprising: a nucleic acid sequence encoding a functional nucleoside diphosphate kinase A (NDPK-A), the nucleic acid sequence being operably linked to regulatory elements that allow for expression of the nucleic acid sequence encoding a functional NDPK-A; (iii) a third genetic entity, comprising: the nucleic acid sequence of interest being operably linked to regulatory elements that allow for expression of the nucleic acid sequence of interest; and (iv) a four genetic entity, comprising: the Epstein Barr virus OriP sequence or one or more subsequences thereof, wherein the one or more subsequences comprise at least the ‘Family of Repeats’ DNA-binding site for EBNA-1 and the ‘Dyad Symmetry’ DNA-binding site for EBNA-1. The present invention also relates to corresponding mammalian host cells and methods for expressing a nucleic acid sequence of interest by means of such expression system.

    Ligand screening and discovery
    6.
    发明授权
    Ligand screening and discovery 有权
    配体筛选和发现

    公开(公告)号：US09562902B2

    公开(公告)日：2017-02-07

    申请号：US14498340

    申请日：2014-09-26

    申请人： Dyax Corp.

    发明人： Hendricus Renerus Jacobus Mattheus Hoogenboom ,  Jurgen Mullberg ,  Robert Charles Ladner

    IPC分类号： C40B30/04 ,  G01N33/569 ,  C07K16/00 ,  C12N15/85 ,  C12N15/10 ,  C12N15/62

    CPC分类号： C12N15/1082 ,  C07K16/00 ,  C07K2317/56 ,  C12N15/1058 ,  C12N15/625 ,  C12N15/85 ,  C12N2800/108 ,  C12N2830/002 ,  C12N2830/50 ,  C12N2830/55 ,  C12N2830/85 ,  C12N2840/203 ,  C40B30/04 ,  G01N33/56983

    摘要： Disclosed is a method that includes: (i) providing a plurality of initial nucleic acid cassettes that include: a) a first coding region encoding a first immunoglobulin variable domain, b) a second coding region encoding a second immunoglobulin variable domain, and c) a ribosomal binding site disposed between the first and second coding regions for translation of the second polypeptide in a first expression system, wherein the first and second coding regions are in the same translational orientation; (ii) modifying each nucleic acid cassette of the plurality in a single reaction mixture so that it is functional in a second expression system, wherein the first and second region remain physically attached during the modifying; (iii) introducing each modified nucleic acid cassette into a mammalian cell to produce a mixture of transfected cells; and (iv) expressing each modified nucleic acid cassette in the transfected cells.

    摘要翻译： 公开了一种方法，其包括：（i）提供多个初始核酸盒，其包括：a）编码第一免疫球蛋白可变结构域的第一编码区，b）编码第二免疫球蛋白可变结构域的第二编码区，和c） 布置在第一和第二编码区之间用于在第一表达系统中翻译第二多肽的核糖体结合位点，其中第一和第二编码区具有相同的平移方向; （ii）在单个反应混合物中修饰多个的每个核酸盒，使得其在第二表达系统中起作用，其中第一和第二区域在修饰期间保持物理附着; （iii）将每个修饰的核酸盒引入哺乳动物细胞以产生转染细胞的混合物; 和（iv）在转染的细胞中表达每个修饰的核酸盒。

    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLERECOGNITION SITES
    9.
    发明申请
    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLERECOGNITION SITES 有权
    使用多重分解位点合成核酸分子的方法和组合物

    公开(公告)号：US20150225729A1

    公开(公告)日：2015-08-13

    申请号：US14578336

    申请日：2014-12-19

    申请人： LIFE TECHNOLOGIES CORPORATION

    发明人： Jonathan CHESNUT ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gary Temple

    IPC分类号： C12N15/66 ,  C12N9/90

    CPC分类号： C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

    摘要： The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

    摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供了包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。