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    PROTEIN EXPRESSION FROM MULTIPLE NUCLEIC ACIDS
    1.
    发明申请
    PROTEIN EXPRESSION FROM MULTIPLE NUCLEIC ACIDS 有权
    来自多种核酸的蛋白质表达

    公开(公告)号：US20100249379A1

    公开(公告)日：2010-09-30

    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： C07K16/28 ,  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试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试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。

    IMMUNOGLOBULIN PRODUCTION
    2.
    发明申请
    IMMUNOGLOBULIN PRODUCTION 有权
    免疫球蛋白生产

    公开(公告)号：US20100184144A1

    公开(公告)日：2010-07-22

    申请号：US12664401

    申请日：2008-06-25

    申请人： Ulrich Goepfert ,  Silke Hansen ,  Hendrik Knoetgen ,  Erhard Kopetzki ,  Oliver Ploettner

    发明人： Ulrich Goepfert ,  Silke Hansen ,  Hendrik Knoetgen ,  Erhard Kopetzki ,  Oliver Ploettner

    IPC分类号： C12P21/00 ,  C07H21/00 ,  C12N15/74 ,  C12N5/10

    CPC分类号： C07K16/00 ,  C07K2317/21 ,  C07K2317/52 ,  C12N2830/42

    摘要： The current invention comprises a nucleic acid encoding the amino acid sequence of the C-terminal part of the CH3-domain of an immunoglobulin of the class IgA or IgG, or the C-terminal part of the CH4-domain of an immunoglobulin of the class IgE or IgM, wherein the glycine-lysine-dipeptide comprised in the amino acid sequence of the C-terminal part of the CH3- or CH4-domain is encoded by the nucleic acid ggaaaa, or the nucleic acid ggcaaa, or the nucleic acid gggaaa, or the nucleic acid gggaag, or the nucleic acid ggcaag, or the nucleic acid ggaaag.

    摘要翻译： 本发明包括编码IgA或IgG类免疫球蛋白的CH3结构域的C末端部分的氨基酸序列的核酸，或类别的免疫球蛋白的CH4结构域的C末端部分 IgE或IgM，其中包含在CH 3 - 或CH 4结构域的C-末端部分的氨基酸序列中的甘氨酸 - 赖氨酸二肽由核酸ggaaaa或核酸ggcaaa或核酸gggaaa ，或核酸gggaag，或核酸ggcaag，或核酸ggaaag。

    Protein Expression in Rodent Cells
    3.
    发明申请
    Protein Expression in Rodent Cells 有权
    啮齿动物细胞中的蛋白质表达

    公开(公告)号：US20100021965A1

    公开(公告)日：2010-01-28

    申请号：US12083535

    申请日：2006-10-26

    申请人： Ulrich Goepfert ,  Erhard Kopetzki

    发明人： Ulrich Goepfert ,  Erhard Kopetzki

    IPC分类号： C12P21/00 ,  C12N15/87 ,  C12N5/18

    CPC分类号： C12N15/85 ,  C12N7/00 ,  C12N2710/16252 ,  C12N2800/108

    摘要： A method is described for the expression of heterologous polypeptides in rodent cells. The method comprises the oriP/EBNA-1 episomal replication and maintenance system of the Epstein-Barr-Virus (EBV). With the stable integration of an EBNA-1-protein expression cassette under the control of a promoter into the genome of a rodent cell an EBNA-1-protein expression in the cells was obtained. The heterologous protein is expressed from an episome comprising an EBV origin of replication and a functional expression cassette of said heterologous protein. The invention further comprises transformed rodent cell lines, a method for the production of a heterologous protein in said cell lines and a kit for the construction of said cell lines.

    摘要翻译： 描述了在啮齿动物细胞中表达异源多肽的方法。 该方法包括爱泼斯坦 - 巴尔病毒（EBV）的oriP / EBNA-1附加体复制和维持系统。 通过将启动子控制下的EBNA-1蛋白表达盒稳定整合到啮齿动物细胞的基因组中，获得细胞中的EBNA-1蛋白表达。 异源蛋白质由包含EBV复制起点的所述异源蛋白质的功能性表达盒的表达来表达。 本发明还包括转化的啮齿动物细胞系，用于在所述细胞系中产生异源蛋白质的方法和用于构建所述细胞系的试剂盒。

    Protein expression from multiple nucleic acids
    4.
    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。

    Heavy chain mutant leading to improved immunoglobulin production
    5.
    发明授权
    Heavy chain mutant leading to improved immunoglobulin production 有权
    重链突变体导致免疫球蛋白产生的改善

    公开(公告)号：US08314225B2

    公开(公告)日：2012-11-20

    申请号：US12664401

    申请日：2008-06-25

    申请人： Ulrich Goepfert ,  Silke Hansen ,  Hendrik Knoetgen ,  Erhard Kopetzki ,  Oliver Ploettner

    发明人： Ulrich Goepfert ,  Silke Hansen ,  Hendrik Knoetgen ,  Erhard Kopetzki ,  Oliver Ploettner

    IPC分类号： C07H21/04

    CPC分类号： C07K16/00 ,  C07K2317/21 ,  C07K2317/52 ,  C12N2830/42

    摘要： The current invention comprises a nucleic acid encoding the amino acid sequence of the C-terminal part of the CH3-domain of an immunoglobulin of the class IgA or IgG, or the C-terminal part of the CH4-domain of an immunoglobulin of the class IgE or IgM, wherein the glycine-lysine-dipeptide comprised in the amino acid sequence of the C-terminal part of the CH3- or CH4-domain is encoded by the nucleic acid ggaaaa, or the nucleic acid ggcaaa, or the nucleic acid gggaaa, or the nucleic acid gggaag, or the nucleic acid ggcaag, or the nucleic acid ggaaag.

    摘要翻译： 本发明包括编码IgA或IgG类免疫球蛋白的CH3结构域的C末端部分的氨基酸序列的核酸，或类别的免疫球蛋白的CH4结构域的C末端部分 IgE或IgM，其中包含在CH 3 - 或CH 4结构域的C-末端部分的氨基酸序列中的甘氨酸 - 赖氨酸二肽由核酸ggaaaa或核酸ggcaaa或核酸gggaaa ，或核酸gggaag，或核酸ggcaag，或核酸ggaaag。

    Protein expression in rodent cells
    6.
    发明授权
    Protein expression in rodent cells 有权
    啮齿动物细胞中的蛋白质表达

    公开(公告)号：US07892790B2

    公开(公告)日：2011-02-22

    申请号：US12083535

    申请日：2006-10-26

    申请人： Ulrich Goepfert ,  Erhard Kopetzki

    发明人： Ulrich Goepfert ,  Erhard Kopetzki

    IPC分类号： C12P21/06 ,  C12N5/10

    CPC分类号： C12N15/85 ,  C12N7/00 ,  C12N2710/16252 ,  C12N2800/108

    摘要： A method is described for the expression of heterologous polypeptides in rodent cells. The method comprises the oriP/EBNA-1 episomal replication and maintenance system of the Epstein-Barr-Virus (EBV). With the stable integration of an EBNA-1-protein expression cassette under the control of a promoter into the genome of a rodent cell an EBNA-1-protein expression in the cells was obtained. The heterologous protein is expressed from an episome comprising an EBV origin of replication and a functional expression cassette of said heterologous protein. The invention further comprises transformed rodent cell lines, a method for the production of a heterologous protein in said cell lines and a kit for the construction of said cell lines.

    摘要翻译： 描述了在啮齿动物细胞中表达异源多肽的方法。 该方法包括爱泼斯坦 - 巴尔病毒（EBV）的oriP / EBNA-1附加体复制和维持系统。 通过将启动子控制下的EBNA-1蛋白表达盒稳定整合到啮齿动物细胞的基因组中，获得细胞中的EBNA-1蛋白表达。 异源蛋白质由包含EBV复制起点的所述异源蛋白质的功能性表达盒的表达来表达。 本发明还包括转化的啮齿动物细胞系，用于在所述细胞系中产生异源蛋白质的方法和用于构建所述细胞系的试剂盒。

    Method for the recombinant expression of a polypeptide
    10.
    发明申请
    Method for the recombinant expression of a polypeptide 有权
    重组表达多肽的方法

    公开(公告)号：US20070117185A1

    公开(公告)日：2007-05-24

    申请号：US11583573

    申请日：2006-10-19

    申请人： Erhard Kopetzki

    发明人： Erhard Kopetzki

    IPC分类号： C12P21/08 ,  C07K16/18 ,  C12P21/06 ,  C07H21/04 ,  C12N5/06

    CPC分类号： C07K14/005 ,  A61K38/162 ,  C07K16/00 ,  C07K16/2863 ,  C07K2317/50 ,  C07K2319/00 ,  C12N2740/16122

    摘要： A method for the recombinant production of a heterologous polypeptide in a eukaryotic host cell is described. The host cell comprises an expression plasmid, whereby the expression plasmid comprises in a 5′ to 3′ direction a) a promoter, b) a nucleic acid encoding a first polypeptide, whose amino acid sequence is selected from Table 1 depending on the first two amino acids of the second polypeptide, c) a nucleic acid encoding a second polypeptide comprising a nucleic acid encoding a heterologous polypeptide, a nucleic acid encoding a linker, and a nucleic acid encoding an immunoglobulin fragment, and d) a 3′ untranslated region comprising a polyadenylation signal. Further a plasmid and a kit are described.

    摘要翻译： 描述了在真核宿主细胞中重组产生异源多肽的方法。 宿主细胞包含表达质粒，其中表达质粒包含5'至3'方向a）启动子，b）编码第一多肽的核酸，其氨基酸序列选自表1，取决于前两个 第二多肽的氨基酸，c）编码包含编码异源多肽的核酸，编码接头的核酸和编码免疫球蛋白片段的核酸的第二多肽的核酸，以及d）3'非翻译区，其包含 多聚腺苷酸化信号。 此外，还描述了质粒和试剂盒。