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    N-Acetylglucosaminyltransferase III Expression in Lower Eukaryotes
    11.
    发明申请
    N-Acetylglucosaminyltransferase III Expression in Lower Eukaryotes 审中-公开
    N-乙酰葡糖胺基转移酶III在低等真核生物中的表达

    公开(公告)号：US20100016555A1

    公开(公告)日：2010-01-21

    申请号：US12540915

    申请日：2009-08-13

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

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

    IPC: C07K16/00 ,  C07K14/435

    CPC classification number: C07K14/39 ,  A61K38/00 ,  C07K2319/05 ,  C12N9/1051 ,  C12P21/005 ,  Y02A50/396

    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 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 activity, which produce bisected N-glycan structures and may 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.

    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。 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活性，其产生二等分的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，糖转运蛋白和甘露糖苷酶）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。

    Production of modified glycoproteins having multiple antennary structures
    12.
    发明申请
    Production of modified glycoproteins having multiple antennary structures 审中-公开
    具有多个触角结构的修饰的糖蛋白的生产

    公开(公告)号：US20090226959A1

    公开(公告)日：2009-09-10

    申请号：US12313636

    申请日：2008-11-21

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

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

    IPC: C12P21/00 ,  C12N1/19 ,  C12N1/15 ,  C12N15/63 ,  C07K2/00

    CPC classification number: C12P21/005 ,  C12N9/1051

    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。

    Method to engineer mammanlian-type carbohydrate structures
    14.
    发明申请
    Method to engineer mammanlian-type carbohydrate structures 有权
    制造马姆兰型碳水化合物结构的方法

    公开(公告)号：US20050170452A1

    公开(公告)日：2005-08-04

    申请号：US10500240

    申请日：2002-12-24

    Applicant: Stefan Wildt ,  Robert Miele ,  Juergen Nett ,  Robert Davidson

    Inventor： Stefan Wildt ,  Robert Miele ,  Juergen Nett ,  Robert Davidson

    IPC: C12N15/09 ,  C07H21/04 ,  C07K14/47 ,  C07K16/00 ,  C12N1/15 ,  C12N1/16 ,  C12N1/18 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N9/10 ,  C12N15/74 ,  C12P21/00 ,  C12P21/02

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

    Abstract: 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.

    Abstract translation: 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GlcNAcMan 3 GlcNAc 2核心结构，然后可以通过异源表达一种或多种酶，例如糖基 - 转运酶，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。

    RECOMBINANT ECTODOMAIN EXPRESSION OF HERPES SIMPLEX VIRUS GLYCOPROTEINS IN YEAST
    17.
    发明申请
    RECOMBINANT ECTODOMAIN EXPRESSION OF HERPES SIMPLEX VIRUS GLYCOPROTEINS IN YEAST 审中-公开
    HERPES的重组ECTODOMAIN表达简单病毒糖蛋白在YEAST

    公开(公告)号：US20120121630A1

    公开(公告)日：2012-05-17

    申请号：US13386508

    申请日：2010-07-21

    Applicant: Janine T. Bryan ,  John W. Ballet ,  Jessica A. Flynn ,  Danilo R. Casimiro ,  Robert C. Davidson ,  Victoria Copeland ,  Sandra E. Rios ,  Byung-Kwon Choi ,  Stefan Wildt

    Inventor： Janine T. Bryan ,  John W. Ballet ,  Jessica A. Flynn ,  Danilo R. Casimiro ,  Robert C. Davidson ,  Victoria Copeland ,  Sandra E. Rios ,  Byung-Kwon Choi ,  Stefan Wildt

    IPC: A61K39/245 ,  A61P37/06 ,  C12N15/38 ,  C07K14/035 ,  C12P21/00

    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-糖型的单纯疱疹病毒（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，其可以进行糖工程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。 重组糖蛋白通过一种方法产生，在一个实施方案中，该方法包括用含有编码HSV糖蛋白的DNA的表达载体转化毕赤酵母属的酵母，所述HSV糖蛋白在毕赤酵母属酵母中起促进作用的启动子的调控， 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。

    SURFACE DISPLAY OF RECOMBINANT PROTEINS IN LOWER EUKARYOTES
    20.
    发明申请
    SURFACE DISPLAY OF RECOMBINANT PROTEINS IN LOWER EUKARYOTES 有权
    重组蛋白的表面显示在较低的毒素中

    公开(公告)号：US20100331192A1

    公开(公告)日：2010-12-30

    申请号：US12863911

    申请日：2009-02-20

    Applicant: Dongxing Zha ,  Stefan Wildt

    Inventor： Dongxing Zha ,  Stefan Wildt

    IPC: C40B10/00 ,  G01N33/53

    CPC classification number: C12N15/1037 ,  C07K16/005 ,  C07K16/2887 ,  C07K16/32 ,  C07K16/40 ,  C07K2317/51 ,  C07K2317/55 ,  C12N15/1055 ,  C40B40/02 ,  C40B40/08 ,  C40B50/06

    Abstract: Methods for display of recombinant proteins or protein libraries on the surface of lower eukaryotes such as yeast and filamentous fungi are described. The methods are useful for screening libraries of recombinant proteins in lower eukaryotes to identify particular proteins with desired properties from the array of proteins in the libraries. The methods are particularly useful for constructing and screening antibody libraries in lower eukaryotes.

    Abstract translation: 描述了在低等真核生物如酵母和丝状真菌的表面上显示重组蛋白或蛋白质文库的方法。 该方法可用于筛选低等真核生物中的重组蛋白的文库，以从文库中的蛋白质阵列鉴定具有所需性质的特定蛋白质。 该方法对于构建和筛选低等真核生物中的抗体文库特别有用。

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