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    Combinatorial DNA library for producing modified N-glycans in lower eukaryotes
    34.
    发明授权
    Combinatorial DNA library for producing modified N-glycans in lower eukaryotes 有权
    用于在低等真核生物中生产修饰的N-聚糖的组合DNA文库

    公开(公告)号：US07449308B2

    公开(公告)日：2008-11-11

    申请号：US10371877

    申请日：2003-02-20

    申请人： Tillman U. Gerngross ,  Stefan Wildt ,  Byung-Kwon Choi ,  Juergen Hermann Nett ,  Piotr Bobrowicz ,  Stephen R. Hamilton ,  Robert C. Davidson

    发明人： Tillman U. Gerngross ,  Stefan Wildt ,  Byung-Kwon Choi ,  Juergen Hermann Nett ,  Piotr Bobrowicz ,  Stephen R. Hamilton ,  Robert C. Davidson

    IPC分类号： C12N15/00

    CPC分类号： C12P21/005 ,  C07K14/47 ,  C12N9/1051 ,  C12N15/1082 ,  C12N15/79

    摘要： The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man5GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.

    摘要翻译： 本发明涉及具有修饰的寡糖的真核宿主细胞，其可以通过异源表达一组糖基转移酶，糖转运体和甘露糖苷酶进一步修饰，以成为用于产生哺乳动物例如人治疗性糖蛋白的宿主菌株。 本发明提供核酸分子和组合文库，其可用于成功靶向和表达哺乳动物酶活性，例如参与糖基化的真核宿主细胞中的细胞内区室的那些。 该方法提供了可用于表达和靶向参与糖基化的任何所需基因的工程化宿主细胞。 建立或选择具有修饰寡糖的宿主细胞。 在工程化的宿主细胞中制备的N-聚糖具有可以通过异源表达一种或多种酶（例如糖基转移酶）而进一步修饰的Man III / 糖转运蛋白和甘露糖苷酶，以产生人样糖蛋白。 为了生产治疗性蛋白质，该方法可适用于工程化可能获得任何所需糖基化结构的细胞系。