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    PRODUCTION OF GLYCOPROTEINS WITH REDUCED O-GLYCOSYLATION
    5.
    发明申请
    PRODUCTION OF GLYCOPROTEINS WITH REDUCED O-GLYCOSYLATION 有权
    生产具有减少的O-糖苷化的糖蛋白

    公开(公告)号：US20120322101A1

    公开(公告)日：2012-12-20

    申请号：US13530768

    申请日：2012-06-22

    Applicant: Piotr Bobrowicz ,  W. James Cook ,  Warren Kett

    Inventor： Piotr Bobrowicz ,  W. James Cook ,  Warren Kett

    IPC: C12P21/00

    CPC classification number: C07K16/32 ,  C07K2317/41 ,  C12P21/005

    Abstract: A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more α-1,2-mannosidases.

    Abstract translation: 描述了一种用于生产具有减少量的O-连接糖基化的蛋白质组合物的方法。 该方法包括在存在Pmt介导的O-连接糖基化的抑制剂和/或在一种或多种α-1,2-甘露糖苷酶的存在下培养的细胞中产生蛋白质。

    PRODUCTION OF GLYCOPROTEINS WITH REDUCED O-GLYCOSYLATION
    6.
    发明申请
    PRODUCTION OF GLYCOPROTEINS WITH REDUCED O-GLYCOSYLATION 有权
    生产具有减少的O-糖苷化的糖蛋白

    公开(公告)号：US20090170159A1

    公开(公告)日：2009-07-02

    申请号：US12084591

    申请日：2006-11-10

    Applicant: Piotr Bobrowicz ,  W. James Cook ,  Warren Kett

    Inventor： Piotr Bobrowicz ,  W. James Cook ,  Warren Kett

    IPC: C12P21/00

    CPC classification number: C07K16/32 ,  C07K2317/41 ,  C12P21/005

    Abstract: A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more α-1,2-mannosidases.

    Abstract translation: 描述了一种用于生产具有减少量的O-连接糖基化的蛋白质组合物的方法。 该方法包括在存在Pmt介导的O-连接糖基化抑制剂和/或在一种或多种α-1,2-甘露糖苷酶存在下培养的细胞中产生蛋白质。

    VECTORS AND YEAST STRAINS FOR PROTEIN PRODUCTION
    7.
    发明申请
    VECTORS AND YEAST STRAINS FOR PROTEIN PRODUCTION 审中-公开
    用于蛋白质生产的载体和YEAST菌株

    公开(公告)号：US20100311122A1

    公开(公告)日：2010-12-09

    申请号：US12863468

    申请日：2009-02-09

    Applicant: Byung-Kwon Choi ,  Piotr Bobrowicz ,  W. James Cook

    Inventor： Byung-Kwon Choi ,  Piotr Bobrowicz ,  W. James Cook

    IPC: C12P21/00 ,  C12N1/19 ,  C12N9/00

    CPC classification number: C12N9/90 ,  C07K14/4725 ,  C12N9/1051 ,  C12N15/815 ,  C12N15/85 ,  C12N2800/102 ,  C12P21/005

    Abstract: Lower eukaryote host cells in which the function of at least one endogenous gene encoding a chaperone protein, such as a Protein Disulphide Isomerase (PDI), has been reduced or eliminated and at least one mammalian homolog of the chaperone protein is expressed are described. In particular aspects, the host cells further include a deletion or disruption of one or more O-protein mannosyltransferase genes, and/or overexpression of an endogenous or exogenous Ca2+ ATPase. These host cells are useful for producing recombinant glycoproteins in large amounts and for producing recombinant glycoproteins that have reduced O-glycosylation.

    Abstract translation: 描述了其中至少一个编码分子伴侣蛋白（例如蛋白质二硫键异构酶（PDI））的内源基因的功能已经被还原或消除并且伴侣蛋白的至少一个哺乳动物同系物被表达的低等真核生物宿主细胞。 在特定方面，宿主细胞还包括一个或多个O-蛋白质甘露糖基转移酶基因的缺失或破坏，和/或内源或外源Ca2 + ATP酶的过度表达。 这些宿主细胞可用于大量生产重组糖蛋白和用于产生具有降低的O-糖基化的重组糖蛋白。

    VECTORS AND YEAST STRAINS FOR PROTEIN PRODUCTION
    8.
    发明申请
    VECTORS AND YEAST STRAINS FOR PROTEIN PRODUCTION 审中-公开
    用于蛋白质生产的载体和YEAST菌株

    公开(公告)号：US20110312032A1

    公开(公告)日：2011-12-22

    申请号：US13207848

    申请日：2011-08-11

    Applicant: Byung-Kwon Choi ,  Piotr Bobrowicz ,  W. James Cook ,  Elena E. Brevnova

    Inventor： Byung-Kwon Choi ,  Piotr Bobrowicz ,  W. James Cook ,  Elena E. Brevnova

    IPC: C12P21/00 ,  C12N9/00 ,  C12N1/19

    CPC classification number: C12N9/90 ,  C07K14/4725 ,  C12N9/1051 ,  C12N15/815 ,  C12N15/85 ,  C12N2800/102 ,  C12P21/005

    Abstract: Lower eukaryote host cells in which the function of at least one endogenous gene encoding a chaperone protein, such as a Protein Disulphide Isomerase (PDI), has been reduced or eliminated and at least one mammalian homolog of the chaperone protein is expressed are described. In particular aspects, the host cells further include a deletion or disruption of one or more O-protein mannosyltransferase genes, and/or overexpression of an endogenous or exogenous Ca2+ATPase. These host cells are useful for producing recombinant glycoproteins in large amounts and for producing recombinant glycoproteins that have reduced O-glycosylation.

    Abstract translation: 描述了其中至少一个编码分子伴侣蛋白（例如蛋白质二硫键异构酶（PDI））的内源基因的功能已经被还原或消除并且伴侣蛋白的至少一个哺乳动物同系物被表达的低等真核生物宿主细胞。 在特定方面，宿主细胞还包括一个或多个O-蛋白质甘露糖基转移酶基因的缺失或破坏，和/或内源或外源Ca2 + ATP酶的过度表达。 这些宿主细胞可用于大量生产重组糖蛋白和用于产生具有降低的O-糖基化的重组糖蛋白。

    Combinatorial DNA Library for Producing Modified N-Glycans In Lower Eukaryotes
    9.
    发明申请
    Combinatorial DNA Library for Producing Modified N-Glycans In Lower Eukaryotes 审中-公开
    用于在低等真核生物中生产修饰的N-糖的组合DNA文库

    公开(公告)号：US20130217067A1

    公开(公告)日：2013-08-22

    申请号：US13408432

    申请日：2012-02-29

    Applicant: TILLMAN U. GERNGROSS ,  Stefan Wildt ,  Byung-kwon Choi ,  Juergen Hermann Nett ,  Piotr Bobrowicz ,  Stephen R. Hamilton ,  Robert C. Davidson

    Inventor： TILLMAN U. GERNGROSS ,  Stefan Wildt ,  Byung-kwon Choi ,  Juergen Hermann Nett ,  Piotr Bobrowicz ,  Stephen R. Hamilton ,  Robert C. Davidson

    IPC: C07K14/47

    CPC classification number: C12P21/005 ,  C07K14/47 ,  C12N9/1051 ,  C12N15/1082 ,  C12N15/79

    Abstract: 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.

    Abstract translation: 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。

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