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    S99T C-11 Truncated polynucleotides encoding interferon gamma polypeptide variants
    1.
    发明授权
    S99T C-11 Truncated polynucleotides encoding interferon gamma polypeptide variants 失效
    S99T C-11编码干扰素γ多肽变体的截短多核苷酸

    公开(公告)号：US07390638B2

    公开(公告)日：2008-06-24

    申请号：US11115906

    申请日：2005-04-27

    申请人： Bart Van Den Hazel ,  Anne Dam Jensen ,  Frank Bech Nygaard ,  Kim Vilbour Andersen

    发明人： Bart Van Den Hazel ,  Anne Dam Jensen ,  Frank Bech Nygaard ,  Kim Vilbour Andersen

    IPC分类号： C12N15/21 ,  C12N5/00 ,  C12N15/00 ,  C07H21/04

    CPC分类号： C07K14/57 ,  A61K38/00

    摘要： When interferon gamma (IFNG) is produced in mammalian cell lines a heterogenous population of IFNG polypeptides is obtained due to C-terminal processing of the IFNG polypeptide. Clearly, this constitutes a severe problem in that valuable polypeptide material is lost and, further, it is necessary to carry out time-consuming and cumbersome purification in order to obtain a homogenous population of active IFNG polypeptides having the desired length. It has now been found that an IFNG fragment containing 132 amino acid residues (truncated at the nucleotide level by introducing a stop-codon after the codon encoding amino acid residue no. 132) does not undergo C-terminal truncation or, at least, is not significantly C-terminally truncated. Furthermore, as the IFNG fragment containing 132 amino acid residues is active, this opens up the possibility of producing a homogenous active IFNG polypeptide in eukaryotic host cells, such as CHO cells. More particularly, the present invention relates to an IFNG polypeptide variant exhibiting IFNG activity and having the amino acid sequence shown in SEQ ID NO:12. In a highly preferred embodiment of the invention, the variant comprises at least one further modification, such as 1-10 further modifications, relative to the amino acid sequence shown in SEQ ID NO:12. A particular preferred further modification is E38N+S40T.

    摘要翻译： 当在哺乳动物细胞系中产生干扰素γ（IFNG）时，由于IFNG多肽的C末端加工，获得IFNG多肽的异源群体。 显然，这构成了一个严重的问题，即有价值的多肽材料丢失，而且还需要进行耗时且繁琐的纯化，以获得具有所需长度的活性IFNG多肽的均质群体。 现在已经发现，含有132个氨基酸残基的IFNG片段（在编码氨基酸残基132号的密码子后通过引入终止密码子在核苷酸水平被截短）不经历C-末端截短，或至少是 不显着C末端截断。 此外，由于含有132个氨基酸残基的IFNG片段是有活性的，因此可以在真核宿主细胞如CHO细胞中产生均一的活性IFNG多肽。 更具体地，本发明涉及表现出IFNG活性且具有SEQ ID NO：12所示氨基酸序列的IFNG多肽变体。 在本发明的一个非常优选的实施方案中，相对于SEQ ID NO：12所示的氨基酸序列，变体包含至少一个其它修饰，例如1-10个其它修饰。 特别优选的进一步修改是E38N + S40T。

    Interferon gamma polypeptide variants
    2.
    发明授权
    Interferon gamma polypeptide variants 失效
    干扰素γ多肽变体

    公开(公告)号：US06958388B2

    公开(公告)日：2005-10-25

    申请号：US10195707

    申请日：2002-07-15

    申请人： Bart Van Den Hazel ,  Anne Dam Jensen ,  Frank Bech. Nygaard ,  Kim Vilbour Andersen

    发明人： Bart Van Den Hazel ,  Anne Dam Jensen ,  Frank Bech. Nygaard ,  Kim Vilbour Andersen

    IPC分类号： A61K38/00 ,  C07K14/57 ,  C07K17/00 ,  A61K38/21 ,  C12P21/04

    CPC分类号： C07K14/57 ,  A61K38/00

    摘要： When interferon gamma (IFNG) is produced in mammalian cell lines a heterogenous population of IFNG polypeptides is obtained due to C-terminal processing of the IFNG polypeptide. Clearly, this constitutes a severe problem in that valuable polypeptide material is lost and, further, it is necessary to carry out time-consuming and cumbersome purification in order to obtain a homogenous population of active IFNG polypeptides having the desired length. It has now been found that an IFNG fragment containing 132 amino acid residues (truncated at the nucleotide level by introducing a stop-codon after the codon encoding amino acid residue no. 132) does not undergo C-terminal truncation or, at least, is not significantly C-terminally truncated. Furthermore, as the IFNG fragment containing 132 amino acid residues is active, this opens up the possibility of producing a homogenous active IFNG polypeptide in eukaryotic host cells, such as CHO cells. More particularly, the present invention relates to an IFNG polypeptide variant exhibiting IFNG activity and having the amino acid sequence shown in SEQ ID NO:12. In a highly preferred embodiment of the invention, the variant comprises at least one further modification, such as 1-10 further modifications, relative to the amino acid sequence shown in SEQ ID NO:12. A particular preferred further modification is E38N+S40T.

    摘要翻译： 当在哺乳动物细胞系中产生干扰素γ（IFNG）时，由于IFNG多肽的C末端加工，获得IFNG多肽的异源群体。 显然，这构成了一个严重的问题，即有价值的多肽材料丢失，而且还需要进行耗时且繁琐的纯化，以获得具有所需长度的活性IFNG多肽的均质群体。 现在已经发现，含有132个氨基酸残基的IFNG片段（在编码氨基酸残基132号的密码子后通过引入终止密码子在核苷酸水平被截短）不经历C-末端截短，或至少是 不显着C末端截断。 此外，由于含有132个氨基酸残基的IFNG片段是有活性的，因此可以在真核宿主细胞如CHO细胞中产生均一的活性IFNG多肽。 更具体地，本发明涉及表现出IFNG活性且具有SEQ ID NO：12所示氨基酸序列的IFNG多肽变体。 在本发明的一个非常优选的实施方案中，相对于SEQ ID NO：12所示的氨基酸序列，变体包含至少一个其它修饰，例如1-10个其它修饰。 特别优选的进一步修改是E38N + S40T。