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    Processes and compositions for the isolation of human relaxin
    6.
    发明授权
    Processes and compositions for the isolation of human relaxin 失效
    用于分离人类松弛素的方法和组合物

    公开(公告)号：US5464756A

    公开(公告)日：1995-11-07

    申请号：US908766

    申请日：1992-07-01

    申请人： Dennis J. Henner ,  Richard L. Vandlen ,  James A. Wilkins ,  Daniel G. Yansura

    发明人： Dennis J. Henner ,  Richard L. Vandlen ,  James A. Wilkins ,  Daniel G. Yansura

    IPC分类号： A61K38/00 ,  A61P13/02 ,  A61P15/00 ,  C07H21/04 ,  C07K1/113 ,  C07K1/12 ,  C07K14/00 ,  C07K14/575 ,  C07K14/64 ,  C12N1/21 ,  C12N15/09 ,  C12N15/12 ,  C12N15/16 ,  C12P21/00 ,  C12P21/02 ,  C12P21/06 ,  C12R1/19 ,  C12N15/70

    CPC分类号： C07K14/64 ,  Y10S435/849 ,  Y10S930/31

    摘要： A process is provided for cleaving a polypeptide into at least two polypeptide components comprising treating a reduced, free-cysteine form of the polypeptide with a cleaving agent under conditions for cleaving the polypeptide at a desired junction between the polypeptide cleavage products. More preferably, the process for cleaving comprises culturing cells containing DNA encoding said polypeptide, wherein at least one Asp codon is present in said DNA at a desired junction between the components to be cleaved from each other, said culturing resulting in expression of the DNA to produce the polypeptide in the host cell culture; and treating a reduced, free-cysteine form of the polypeptide with dilute acid under conditions for cleaving the polypeptide at the Asp junction. In particular embodiments, a DNA sequence is provided that encodes a relaxin precursor and includes codons encoding aspartic acid-containing linkers at novel positions within the precursor, allowing the ready cleavage of relaxin A peptides by treatment with dilute acid.

    摘要翻译： 提供了一种用于将多肽切割成至少两种多肽组分的方法，包括在多肽切割产物之间的所需连接处切割多肽的条件下用切割剂处理还原的游离半胱氨酸形式的多肽。 更优选地，所述切割方法包括培养含有编码所述多肽的DNA的细胞，其中至少一个Asp密码子存在于所述DNA中在待切割的组分之间的所需连接处，所述培养导致DNA的表达 在宿主细胞培养物中产生多肽; 以及在Asp连接处切割多肽的条件下用稀酸处理还原的游离半胱氨酸形式的多肽。 在具体实施方案中，提供了编码松弛素前体的DNA序列，并且包括在前体内的新位置编码含有天冬氨酸的接头的密码子，从而允许通过用稀酸处理来快速切割松弛素A肽。

    Process for producing heterologous polypeptides
    7.
    发明授权
    Process for producing heterologous polypeptides 失效
    用于产生异源多肽的方法

    公开(公告)号：US5411873A

    公开(公告)日：1995-05-02

    申请号：US928697

    申请日：1992-08-11

    申请人： Robin M. Adams ,  Scott D. Power ,  David B. Powers ,  James A. Wells ,  Daniel G. Yansura

    发明人： Robin M. Adams ,  Scott D. Power ,  David B. Powers ,  James A. Wells ,  Daniel G. Yansura

    IPC分类号： C11D3/386 ,  C12N9/54 ,  C12N15/10 ,  C12N15/52 ,  C12N15/75 ,  C12P21/02

    CPC分类号： C12N15/52 ,  C11D3/386 ,  C12N15/102 ,  C12N15/75 ,  C12N9/54 ,  C12R1/07

    摘要： Processes for producing various heterologous polypeptides which when expressed are either incorrectly processed and hence asssociated with the surface of the host cell or are not processed to mature form. More specifically, processes for the production of heterologous non-human carbonyl hydrolases expressed either in host cells incapable of producing enzymatically active endoprotease or host cells deficient in enzymatically active extracellular endoprotease are disclosed. Such non-human carbonyl hydrolases generally are incapable of autoproteolytic maturation and become associated with the surface of expression hosts which are deficient in enzymatically active extracellular endoprotease. Processes for preparing non-human carbonyl hydrolase and heterologous polypeptides which are expressed as part of a fusion polypeptide are also disclosed, as well as non-human carbonyl hydrolases which are substantially free of the host cell membrane with which they are normally associated.

    摘要翻译： 用于产生各种异源多肽的方法，当被表达时，它们被错误地加工并因此与宿主细胞的表面相结合或者不被加工成成熟形式。 更具体地，公开了在不能产生酶促活性内切蛋白酶的宿主细胞中表达的异源非人羰基水解酶的生产方法或在酶活性细胞外内蛋白酶中缺乏的宿主细胞。 这种非人羰基水解酶通常不能自身蛋白水解成熟，并且与酶活性细胞外内蛋白酶缺陷的表达宿主的表面相关联。 还公开了制备非人羰基水解酶和作为融合多肽的一部分表达的异源多肽的方法，以及基本上不含与它们通常结合的宿主细胞膜的非人羰基水解酶。