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    • 1. 发明申请
    • TRIPLE VECTOR FOR EXPRESSING ANTIBODY MOLECULES IN FULL THERAPEUTIC FORMAT
    • 用于在完整治疗形式中表达抗体分子的三重载体
    • WO2018069416A1
    • 2018-04-19
    • PCT/EP2017/075983
    • 2017-10-11
    • ARGENX BVBAFAIRJOURNEY BIOLOGICS
    • PAJUELO, Maria GonzalesDE HAARD, Johannes Joseph Wilhelmus
    • C12N15/63
    • C07K16/005C07K2317/52C07K2317/524C07K2317/526C07K2317/53C07K2317/569C12N15/63C12N15/86C12N2830/00C12N2840/00
    • A triple expression vector is disclosed for expressing an antibody molecule comprising an Fc domain in prokaryotic cells and in eukaryotic cells. The triple expression vector comprises a polynucleotide encoding an Fc domain; a polynucleotide encoding a phage coat protein; a cloning site for cloning genes encoding an antibody molecule or a part thereof wherein the antibody molecule or part thereof does not comprise an Fc domain; a prokaryotic secretion signal sequence and a eukaryotic secretion signal sequence, or a secretion signal sequence that drives efficient secretion in both prokaryotic and eukaryotic cells; a promoter for mediating expression in eukaryotic cells; and a stop codon for preventing expression of the phage coat protein in eukaryotic cells. The triple expression vector can be used for expressing an antibody molecule in a phage display format; for producing the antibody molecule in a prokaryotic cell, for example in the periplasm of a prokaryotic cell; and for producing the antibody molecule in a eukaryotic cell, for example a mammalian cell, more particularly a human cell. The antibody molecule contains an Fc domain, and may be for example a VHH-Fc molecule or an scFv-Fc molecule or a VH-Fc or a VL-Fc. Phage display libraries produced with the vector present the antibody molecule in its therapeutic format. Use of the vector avoids the need for repeated cloning when moving from one expression medium to another.
    • 公开了用于在原核细胞和真核细胞中表达包含Fc结构域的抗体分子的三重表达载体。 三重表达载体包含编码Fc结构域的多核苷酸; 编码噬菌体外壳蛋白的多核苷酸; 克隆编码抗体分子或其部分的基因的克隆位点,其中抗体分子或其部分不包含Fc结构域; 原核分泌信号序列和真核分泌信号序列,或驱动原核细胞和真核细胞中有效分泌的分泌信号序列; 用于介导真核细胞中的表达的启动子; 和用于防止真核细胞中噬菌体外壳蛋白表达的终止密码子。 三重表达载体可用于以噬菌体展示形式表达抗体分子; 用于在原核细胞中产生抗体分子,例如在原核细胞的周质中; 并且用于在真核细胞例如哺乳动物细胞,更特别是人细胞中产生抗体分子。 抗体分子含有Fc结构域,并且可以是例如VHH-Fc分子或scFv-Fc分子或VH-Fc或VL-Fc。 用载体产生的噬菌体展示文库以其治疗形式呈递抗体分子。 使用载体避免了从一种表达载体移动到另一种载体时需要重复克隆。