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    • 76. 发明申请
    • Methods for producing modified glycoproteins
    • 生产改性糖蛋白的方法
    • US20070178551A1
    • 2007-08-02
    • US11265444
    • 2005-11-01
    • Tillman Gerngross
    • Tillman Gerngross
    • C12P21/04C12P21/06C07H21/04C12N1/18C07K14/52C07K16/46
    • C12P21/005C07K2319/04C07K2319/05C12N1/14C12N9/1048C12N9/2488C12N15/79C12N15/80C12N15/81C12Y302/01C12Y302/01113
    • Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar, if not substantially identical, to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways. This is achieved using a combination of engineering and/or selection of strains which: do not express certain enzymes which create the undesirable complex structures characteristic of the fungal glycoproteins, which express exogenous enzymes selected either to have optimal activity under the conditions present in the fungi where activity is desired, or which are targeted to an organelle where optimal activity is achieved, and combinations thereof wherein the genetically engineered eukaryote expresses multiple exogenous enzymes required to produce “human-like” glycoproteins.
    • 已经开发了具有遗传修饰的糖基化途径的细胞系,其允许它们进行模拟人类中糖蛋白的加工的酶反应序列。 在这些工程化宿主中表达的重组蛋白可以产生与其对应物更相似的糖蛋白(如果基本上不相同)。 通常产生含有高甘露糖的N-聚糖(包括单细胞和多细胞真菌)的低等真核生物被修饰以产生N-聚糖,例如Man 3或GlcNAc 2 N或其它结构 沿着人类糖基化途径。 这是使用以下工程和/或选择的组合实现的:不表达产生真菌糖蛋白特征的不合需要的复合结构的某些酶,其表达选择在真菌中存在的条件下具有最佳活性的外源酶 其中需要活性或靶向实现最佳活性的细胞器,以及其组合,其中遗传工程真核生物表达产生“人样”糖蛋白所需的多种外源酶。