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。