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    METHOD FOR THE PRODUCTION OF CONJUGATES OF INSULIN-LIKE GROWTH FACTOR-1 AND POLY(ETHYLENE GLYCOL)
    3.
    发明申请
    METHOD FOR THE PRODUCTION OF CONJUGATES OF INSULIN-LIKE GROWTH FACTOR-1 AND POLY(ETHYLENE GLYCOL) 审中-公开
    产生胰岛素样生长因子-1和聚（乙二醇）的方法

    公开(公告)号：US20100035817A1

    公开(公告)日：2010-02-11

    申请号：US12576266

    申请日：2009-10-09

    申请人： Stephan Fischer ,  Friederike Hesse ,  Hendrik Knoetgen ,  Kurt Lang ,  Friedrich Metzger ,  Joerg Thomas Regula ,  Christian Schantz ,  Andreas Schaubmar ,  Hans-Joachim Schoenfeld

    发明人： Stephan Fischer ,  Friederike Hesse ,  Hendrik Knoetgen ,  Kurt Lang ,  Friedrich Metzger ,  Joerg Thomas Regula ,  Christian Schantz ,  Andreas Schaubmar ,  Hans-Joachim Schoenfeld

    IPC分类号： A61K38/18 ,  A61P25/28

    CPC分类号： C07K14/65 ,  A61K47/60 ,  C07K2319/21 ,  C07K2319/50

    摘要： The present invention relates to a fusion protein comprising IGF-I or an IGF-I variant N-terminally linked to the C-terminus of a propeptide. The invention relates also to a method involving the use of the aforementioned fusion protein in the production of a lysine-PEGylated IGF-I or IGF-I variant. The method comprises the steps of cultivating a prokaryotic host cell comprising an expression vector containing a nucleic acid encoding the fusion protein and causing the cell to express the fusion protein, recovering and PEGylating said fusion protein, cleaving said PEGylated fusion protein with IgA protease, and recovering lysine-PEGylated IGF-I or IGF-I variant. The invention relates also to a lysine-PEGylated IGF-I or IGF-I variant produced using the above method. In addition, the invention relates to a method for treating a neurodegenerative disorders like Alzheimer's Disease using the lysine-PEGylated IGF-I or IGF-I variant and a composition comprising the lysine-PEGylated IGF-I or IGF-I variant.

    摘要翻译： 本发明涉及包含IGF-I或与前肽的C-末端N末端连接的IGF-1变体的融合蛋白。 本发明还涉及涉及在制备赖氨酸 - PEG化IGF-I或IGF-I变体中使用上述融合蛋白的方法。 该方法包括以下步骤：培养包含含有编码融合蛋白的核酸的表达载体的原核宿主细胞，并使细胞表达融合蛋白，回收聚乙二醇化所述融合蛋白，用IgA蛋白酶切割所述聚乙二醇化融合蛋白，以及 回收赖氨酸 - PEG化IGF-I或IGF-I变体。 本发明还涉及使用上述方法制备的赖氨酸 - 聚乙二醇化IGF-I或IGF-I变体。 此外，本发明涉及使用赖氨酸PEG化IGF-I或IGF-1变体和包含赖氨酸 - PEG化IGF-I或IGF-I变体的组合物治疗神经变性疾病如阿尔茨海默病的方法。

    Method for the production of conjugates of insulin-like growth factor-1 and poly(ethylene glycol)
    8.
    发明授权
    Method for the production of conjugates of insulin-like growth factor-1 and poly(ethylene glycol) 有权
    生产胰岛素样生长因子-1和聚（乙二醇）共轭物的方法

    公开(公告)号：US07625996B2

    公开(公告)日：2009-12-01

    申请号：US11846857

    申请日：2007-08-29

    申请人： Stephan Fischer ,  Friederike Hesse ,  Hendrik Knoetgen ,  Kurt Lang ,  Friedrich Metzger ,  Joerg Thomas Regula ,  Christian Schantz ,  Andreas Schaubmar ,  Hans-Joachim Schoenfeld

    发明人： Stephan Fischer ,  Friederike Hesse ,  Hendrik Knoetgen ,  Kurt Lang ,  Friedrich Metzger ,  Joerg Thomas Regula ,  Christian Schantz ,  Andreas Schaubmar ,  Hans-Joachim Schoenfeld

    IPC分类号： A61K38/30 ,  A61K38/00

    CPC分类号： C07K14/65 ,  A61K47/60 ,  C07K2319/21 ,  C07K2319/50

    摘要： The present invention relates to a fusion protein comprising IGF-I or an IGF-I variant N-terminally linked to the C-terminus of a propeptide. The invention relates also to a method involving the use of the aforementioned fusion protein in the production of a lysine-PEGylated IGF-I or IGF-I variant. The method comprises the steps of cultivating a prokaryotic host cell comprising an expression vector containing a nucleic acid encoding the fusion protein and causing the cell to express the fusion protein, recovering and PEGylating said fusion protein, cleaving said PEGylated fusion protein with IgA protease, and recovering lysine-PEGylated IGF-I or IGF-I variant. The invention relates also to a lysine-PEGylated IGF-I or IGF-I variant produced using the above method. In addition, the invention relates to a method for treating a neurodegenerative disorders like Alzheimer's Disease using the lysine-PEGylated IGF-I or IGF-I variant and a composition comprising the lysine-PEGylated IGF-I or IGF-I variant.

    摘要翻译： 本发明涉及包含IGF-I或与前肽的C-末端N末端连接的IGF-1变体的融合蛋白。 本发明还涉及涉及在制备赖氨酸 - PEG化IGF-I或IGF-I变体中使用上述融合蛋白的方法。 该方法包括以下步骤：培养包含含有编码融合蛋白的核酸的表达载体的原核宿主细胞，并使细胞表达融合蛋白，回收聚乙二醇化所述融合蛋白，用IgA蛋白酶切割所述聚乙二醇化融合蛋白，以及 回收赖氨酸 - PEG化IGF-I或IGF-I变体。 本发明还涉及使用上述方法制备的赖氨酸 - 聚乙二醇化IGF-I或IGF-I变体。 此外，本发明涉及使用赖氨酸PEG化IGF-I或IGF-1变体和包含赖氨酸 - PEG化IGF-I或IGF-I变体的组合物治疗神经变性疾病如阿尔茨海默病的方法。

    Protein expression from multiple nucleic acids
    9.
    发明授权
    Protein expression from multiple nucleic acids 有权
    来自多个核酸的蛋白质表达

    公开(公告)号：US08771988B2

    公开(公告)日：2014-07-08

    申请号：US12681781

    申请日：2008-10-09

    申请人： Ulrich Goepfert ,  Hendrik Knoetgen ,  Erhard Kopetzki ,  Anne Stern

    发明人： Ulrich Goepfert ,  Hendrik Knoetgen ,  Erhard Kopetzki ,  Anne Stern

    IPC分类号： C12P21/08

    CPC分类号： C12N15/85 ,  C07K16/18 ,  C07K16/28 ,  C07K16/2812 ,  C07K16/2854 ,  C07K16/2866 ,  C07K16/2896 ,  C07K2317/14 ,  C07K2317/21 ,  C07K2317/51 ,  C07K2317/515 ,  C12P21/00

    摘要： The current invention reports a method for the recombinant production of a secreted heterologous immunoglobulin in a CHO cell comprising the following steps: i) providing a CHO cell, which is adapted to growth in suspension culture, adapted to growth in serum-free medium, mycoplasma free, and virus free, ii) providing a vector comprising a prokaryotic origin of replication, a first nucleic acid conferring resistance to a prokaryotic selection agent, a second nucleic acid encoding the heavy chain of said heterologous immunoglobulin, a third nucleic acid encoding the light chain of said heterologous immunoglobulin, a fourth nucleic acid conferring resistance to a eukaryotic selection agent, iii) transfecting said CHO cell, wherein said transfecting comprises a) transfecting said CHO cell with said vector comprising a fourth nucleic acid conferring resistance to a first eukaryotic selection agent, b) selecting a CHO cell by growth in cultivation medium containing said first eukaryotic selection agent, c) transfecting said selected CHO cell with said vector comprising a fourth nucleic acid conferring resistance to a second eukaryotic selection agent different to said first eukaryotic selection agent, d) selecting a CHO cell by selected growth in cultivation medium containing said first and said second eukaryotic selection agent, iv) cultivating said transfected CHO cell in a medium in the presence of said first and second eukaryotic selection agent, under conditions suitable for the expression of said second, and third nucleic acid, and v) recovering said secreted heterologous immunoglobulin from the cultivation medium.

    摘要翻译： 本发明报告了在CHO细胞中重组产生分泌的异源免疫球蛋白的方法，其包括以下步骤：i）提供适于在悬浮培养物中生长的CHO细胞，适于在无血清培养基中生长，支原体 免费和无病毒，ii）提供包含原核复制起点的载体，赋予原核选择剂抗性的第一核酸，编码所述异源免疫球蛋白的重链的第二核酸，编码光的第三核酸 所述异源免疫球蛋白链，赋予对真核选择剂的抗性的第四核酸，iii）转染所述CHO细胞，其中所述转染包括a）用包含第四核酸的所述载体转染所述CHO细胞，所述第四核酸赋予对第一真核选择性的抗性 试剂，b）通过在含有所述第一种真核生物的培养基中生长来选择CHO细胞 c）用所述载体转染所述选择的CHO细胞，所述载体包含赋予与所述第一真核选择剂不同的第二真核选择剂的抗性的第四核酸，d）通过在含有所述第一和第二真核选择剂的培养基中选择生长来选择CHO细胞， 所述第二真核选择剂，iv）在所述第一和第二真核选择剂的存在下，在适于表达所述第二和第三核酸的条件下，在培养基中培养所述转染的CHO细胞，和v）回收所述分泌的异源 免疫球蛋白。