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    Enzymatic method for modification of recombinant polypeptides
    1.
    发明授权
    Enzymatic method for modification of recombinant polypeptides 失效
    用于修饰重组多肽的酶法

    公开(公告)号：US06403361B1

    公开(公告)日：2002-06-11

    申请号：US09505991

    申请日：2000-02-17

    申请人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

    发明人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

    IPC分类号： C12N120

    CPC分类号： C07K14/60 ,  A61K38/00 ,  C07K14/605 ,  C07K2319/00 ,  C12P21/02 ,  C12P21/06

    摘要： The method of the invention provides for the formation of a recombinant polypeptide which has been modified at the C-terminal end through the use of a transpeptidation process. The method is suitable for modifying recombinant polypeptides of any source including those which may be commercially available, those derived from recombinant single copy or multicopy polypeptide constructs, or those derived from single or multicopy recombinant fusion protein constructs. The transpeptidation reaction involves contacting an endopeptidase enzyme with a recombinant polypeptide to substitute an addition unit, of one or more amino acids, for a leaving unit, linked to a core polypeptide through a cleavage site recognized by the endopeptidase enzyme. Recombinant polypeptides derived from multicopy polypeptide constructs may be cleaved from the multicopy polypeptide at the N-terminal and C-terminal ends and simultaneously under go substitution of the leaving unit by the desired addition unit. The invention utilizes known and newly discovered cleavage recognition sites to effectuate the desired modification products.

    摘要翻译： 本发明的方法提供了通过使用转肽酶方法在C末端修饰的重组多肽的形成。 该方法适用于修饰任何来源的重组多肽，包括可商购的重组多肽，衍生自重组单拷贝或多拷贝多肽构建体的重组多肽，或衍生自单拷贝或多拷贝重组融合蛋白构建体的重组多肽。 转肽反应包括使内肽酶与重组多肽接触，以将一个或多个氨基酸的加成单元替换为通过内肽酶识别的切割位点与核心多肽连接的离去单元。 衍生自多拷贝多肽构建体的重组多肽可以在N末端和C-末端的多拷贝多肽中切割，同时在离去单位取代所需的加成单位。 本发明利用已知和新发现的裂解识别位点来实现所需的修饰产物。

    Enzymatic method for modification or recombinant polypeptides
    2.
    发明授权
    Enzymatic method for modification or recombinant polypeptides 失效
    用于修饰或重组多肽的酶法

    公开(公告)号：US5512459A

    公开(公告)日：1996-04-30

    申请号：US95162

    申请日：1993-07-20

    申请人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

    发明人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

    IPC分类号： C12N15/09 ,  A61K38/00 ,  A61K38/04 ,  A61P3/08 ,  A61P13/02 ,  A61P15/00 ,  A61P43/00 ,  C07K14/60 ,  C07K14/605 ,  C12N1/21 ,  C12P21/02 ,  C12P21/06 ,  C12R1/19 ,  C12N9/74 ,  C12N9/76

    CPC分类号： C07K14/60 ,  C07K14/605 ,  C12P21/02 ,  C12P21/06 ,  A61K38/00 ,  C07K2319/00

    摘要： The method of the invention provides for the formation of a recombinant polypeptide which has been modified at the C-terminal end through the use of a transpeptidation process. The method is suitable for modifying recombinant polypeptides of any source including those which may be commercially available, those derived from recombinant single copy or multicopy polypeptide constructs, or those derived from single or multicopy recombinant fusion protein constructs. The transpeptidation reaction involves contacting an endopeptidase enzyme with a recombinant polypeptide to substitute an addition unit, of one or more amino acids, for a leaving unit, linked to a core polypeptide through a cleavage site recognized by the endopeptidase enzyme. Recombinant polypeptides derived from multicopy polypeptide constructs may be cleaved from the multicopy polypeptide at the N-terminal and C-terminal ends and simultaneously under go substitution of the leaving unit by the desired addition unit. The invention utilizes known and newly discovered cleavage recognition sites to effectuate the desired modification products.

    摘要翻译： 本发明的方法提供了通过使用转肽酶方法在C末端修饰的重组多肽的形成。 该方法适用于修饰任何来源的重组多肽，包括可商购的重组多肽，衍生自重组单拷贝或多拷贝多肽构建体的重组多肽，或衍生自单拷贝或多拷贝重组融合蛋白构建体的重组多肽。 转肽反应包括使内肽酶与重组多肽接触，以将一个或多个氨基酸的加成单元替换为通过内肽酶识别的切割位点与核心多肽连接的离去单元。 衍生自多拷贝多肽构建体的重组多肽可以在N末端和C-末端的多拷贝多肽中切割，同时在离去单位取代所需的加成单位。 本发明利用已知和新发现的裂解识别位点来实现所需的修饰产物。