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    PRODUCTION OF MODIFIED GLYCOPROTEINS HAVING MULTIPLE ANTENNARY STRUCTURES
    3.
    发明申请
    PRODUCTION OF MODIFIED GLYCOPROTEINS HAVING MULTIPLE ANTENNARY STRUCTURES 审中-公开
    生产具有多种天然结构的改性糖蛋白

    公开(公告)号：WO2004074461A3

    公开(公告)日：2005-03-17

    申请号：PCT/US2004005191

    申请日：2004-02-20

    Applicant: BOBROWICZ PIOTR ,  HAMILTON STEPHEN R ,  GERNGROSS TILMAN U ,  WILDT STEFAN ,  CHOI BYUNG-KWON ,  NETT JUERGEN HERMANN ,  DAVIDSON ROBERT C

    Inventor： BOBROWICZ PIOTR ,  HAMILTON STEPHEN R ,  GERNGROSS TILMAN U ,  WILDT STEFAN ,  CHOI BYUNG-KWON ,  NETT JUERGEN HERMANN ,  DAVIDSON ROBERT C

    IPC: A61K38/00 ,  C07K14/39 ,  C07K14/47 ,  C12N1/18 ,  C12N9/10 ,  C12N15/12 ,  C12N15/74 ,  C12P21/00 ,  C12P21/06 ,  C12N5/10 ,  C12N15/81

    CPC classification number: C07K14/39 ,  A61K38/00 ,  C07K2319/05 ,  C12N9/1051 ,  C12P21/005

    Abstract: 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.

    Abstract translation: 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。 在工程化的宿主细胞中制备的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。

    RECOMBINANT ECTODOMAIN EXPRESSION OF HERPES SIMPLEX VIRUS GLYCOPROTEINS IN YEAST
    7.
    发明申请
    RECOMBINANT ECTODOMAIN EXPRESSION OF HERPES SIMPLEX VIRUS GLYCOPROTEINS IN YEAST 审中-公开
    HERPES的重组ECTODOMAIN表达在YEAST中的简单病毒糖蛋白

    公开(公告)号：WO2011011495A1

    公开(公告)日：2011-01-27

    申请号：PCT/US2010/042719

    申请日：2010-07-21

    Applicant: MERCK SHARP & DOHME CORP. ,  BRYAN, Janine, T. ,  BALLIET, John, W. ,  FLYNN, Jessica, A. ,  CASIMIRO, Danilo, R. ,  DAVIDSON, Robert, C. ,  COPELAND, Victoria ,  RIOS, Sandra, E. ,  CHOI, Byun-Kwon ,  WILDT, Stefan

    Inventor： BRYAN, Janine, T. ,  BALLIET, John, W. ,  FLYNN, Jessica, A. ,  CASIMIRO, Danilo, R. ,  DAVIDSON, Robert, C. ,  COPELAND, Victoria ,  RIOS, Sandra, E. ,  CHOI, Byun-Kwon ,  WILDT, Stefan

    IPC: A61K39/245 ,  C07K1/00

    CPC classification number: A61K39/245 ,  A61K39/12 ,  A61K2039/55505 ,  A61K2039/55577 ,  A61K2039/57 ,  C12N2710/16622 ,  C12N2710/16634

    Abstract: The present invention provides Herpes Simplex Virus (HSV) gD, gC, gB and/or gE recombinant glycoproteins having a particular pre-selected N-linked glycosylation pattern as the predominant N-glycoform. The present invention also provides methods of producing these recombinant glycoproteins in yeast, preferably Pichia pastoris , which may be glycoengineered to provide particular glycosylation patterns. The present invention further provides vaccines comprising gD and gC, and optionally gB and/or gE, at least one of which has a particular pre-selected N-linked glycosylation pattern as the predominant N-glycoform. The recombinant glycoproteins are produced by a method which, in one embodiment, comprises transforming a yeast of the genus Pichia with an expression vector containing a DNA encoding an HSV glycoprotein, which is under regulation of a promoter functional in a yeast of the genus Pichia, culturing the yeast in a medium, and recovering the recombinant glycoprotein from the obtained culture. DNA encoding the recombinant glycoproteins is preferably codon-optimized to achieve optimal expression in Pichia .

    Abstract translation: 本发明提供具有特定预先选择的N-连接的糖基化模式作为主要N-糖基的单纯疱疹病毒（HSV）gD，gC，gB和/或gE重组糖蛋白。 本发明还提供了在酵母，优选巴斯德毕赤酵母中生产这些重组糖蛋白的方法，其可以糖工程化以提供特定的糖基化模式。 本发明还提供了包含gD和gC以及任选的gB和/或gE的疫苗，其中至少一种具有特定的预先选择的N-连接的糖基化模式作为主要的N-糖基型。 通过一种方法制备重组糖蛋白，该方法在一个实施方案中包括用含有编码HSV糖蛋白的DNA的表达载体转化毕赤酵母属的酵母，所述HSV糖蛋白处于毕赤酵母属酵母中起促进作用的启动子的调节下， 在培养基中培养酵母，并从获得的培养物中回收重组糖蛋白。 编码重组糖蛋白的DNA优选密码子优化以在毕赤酵母中达到最佳表达。

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