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    Combinatorial DNA library for producing modified N-glycans in lower eukaryotes
    3.
    发明申请
    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：US20090209024A1

    公开(公告)日：2009-08-20

    申请号：US12156936

    申请日：2008-06-05

    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： C12N1/19 ,  C12N1/15

    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。 在工程化宿主细胞中制备的N-聚糖具有Man5GlcNAc2核心结构，然后可以通过异源表达一种或多种酶，例如糖基转移酶，糖转运蛋白和甘露糖苷酶来进一步修饰，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。

    Method to engineer mammalian-type carbohydrate structures
    4.
    发明授权
    Method to engineer mammalian-type carbohydrate structures 有权
    哺乳动物型碳水化合物结构的设计方法

    公开(公告)号：US08932825B2

    公开(公告)日：2015-01-13

    申请号：US10500240

    申请日：2002-12-24

    申请人： Stefan Wildt ,  Robert Gordon Miele ,  Juergen Hermann Nett ,  Robert C. Davidson

    发明人： Stefan Wildt ,  Robert Gordon Miele ,  Juergen Hermann Nett ,  Robert C. Davidson

    IPC分类号： G01N33/569 ,  C12P1/02 ,  C12N1/00 ,  A61K51/00 ,  C12N9/10 ,  C12P21/00

    CPC分类号： C12N15/815 ,  A01K2217/075 ,  C07K14/705 ,  C07K2319/05 ,  C12N9/1051 ,  C12N9/2402 ,  C12P21/005 ,  C12Y204/01143 ,  C12Y204/01144 ,  C12Y204/01145 ,  C12Y204/01155 ,  C12Y302/01024

    摘要： The present invention relates to host cells having modified lipid-linked 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 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 lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells have a GlcNAcMan3GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g., glycosyl-transferases, 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： 本发明涉及具有经修饰的脂质连接的寡糖的宿主细胞，其可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。 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。 制备或选择具有修饰的脂质连接寡糖的宿主细胞。 在工程化的宿主细胞中制备的N-聚糖具有GlcNAcMan3GlcNAc2核心结构，然后可以通过异源表达一种或多种酶，例如糖基转移酶，糖转运蛋白和甘露糖苷酶来进一步修饰，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。

    Combinatorial DNA library for producing modified N-glycans in lower eukaryotes
    5.
    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：US08877462B2

    公开(公告)日：2014-11-04

    申请号：US13408432

    申请日：2012-02-29

    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： C12N1/14

    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.

    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。 在工程化宿主细胞中制备的N-聚糖具有Man5GlcNAc2核心结构，然后可以通过异源表达一种或多种酶，例如糖基转移酶，糖转运蛋白和甘露糖苷酶来进一步修饰，以产生人样糖蛋白。

    Production of modified glycoproteins having multiple antennary structures
    6.
    发明授权
    Production of modified glycoproteins having multiple antennary structures 失效
    具有多个触角结构的修饰的糖蛋白的生产

    公开(公告)号：US08697394B2

    公开(公告)日：2014-04-15

    申请号：US10546101

    申请日：2004-02-20

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

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

    IPC分类号： C12P21/02 ,  C12N1/00 ,  C12N1/15 ,  C12N15/00

    CPC分类号： C12P21/005 ,  C12N9/1051

    摘要： The present invention relates to eukaryotic host cells, especially lower eukaryotic host cells, having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar and sugar nucleotide transporters to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. 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 lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells exhibit GnTIII, GnTIV, GnTV, GnT VI or GnTIX activity, which produce bisected and/or multiantennary N-glycan structures and may be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar, sugar nucleotide transporters, 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，特别是较低的真核宿主细胞，其可以通过异源表达一组糖基转移酶，糖和糖核苷酸转运蛋白进一步修饰，以成为用于产生哺乳动物的宿主菌株， 人类治疗糖蛋白。 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。 制备或选择具有修饰的脂质连接寡糖的宿主细胞。 在工程化宿主细胞中制备的N-聚糖表现出GnTIII，GnTIV，GnTV，GnT VI或GnTIX活性，其产生二等分和/或多元N-聚糖结构，并且可以通过异源表达一种或多种酶，例如糖基转移酶 ，糖，糖核苷酸转运蛋白，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。