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12. 发明授权

US5100788A Method of producing and isolating IGG-binding protein a fusion peptides and a vector therefor 失效
标题翻译：生产和分离IGG结合蛋白的融合蛋白及其载体的方法
公开(公告)号：US5100788A
公开(公告)日：1992-03-31
申请号：US196846
申请日：1988-05-09
申请人： Sven Lofdahl , Mathias Uhlen , Martin Lindberg , John Sjoquist
发明人： Sven Lofdahl , Mathias Uhlen , Martin Lindberg , John Sjoquist
IPC分类号： C12N15/00 , A61K35/74 , A61K38/00 , C07H1/00 , C07H21/04 , C07K1/00 , C07K1/14 , C07K1/22 , C07K14/65 , C07K16/00 , C07K17/02 , C07K19/00 , C12N1/20 , C12N15/09 , C12N15/11 , C12N15/62 , C12N15/66 , C12P21/00 , C12P21/02 , C12R1/07 , C12R1/19 , C12R1/44
CPC分类号： C07K14/65 , C12N15/11 , C12N15/62 , C12N15/66 , C12P21/02 , C07K2319/00 , C07K2319/036 , C07K2319/61 , C07K2319/705 , C07K2319/75
摘要： A method of producing and selectively isolating a desired protein or polypeptide or derivative thereof by constructing a recombinant vector comprising a DNA sequence coding for said desired protein or polypeptide operatively linked to a DNA sequence coding for protein A or an active polypeptide fragment thereof or any other macromolecule capable of binding to the constant regions of immunoglobulins, such that said DNA sequences together code for an IgG-binding fusion product between said desired protein or polypeptide and said protein A, active polypeptide fragment thereof or macromolecule; transforming a compatible host with said recombinant vector such that the combined DNA sequences coding for said fusion protein or polypeptide can be expressed by the host, and culturing the transformed host in a suitable growth medium to produce said fusion protein or polypeptide; selectively isolating said fusion protein or polypeptide by adsorption to an IgG-supporting carrier material; and optionally desorbing said fusion protein or polypeptide from said IgG-supporting carrier, said fusion protein or polypeptide coded for by said combined DNA-sequence optionally comprising a unique cleavage site between said protein A part and said desired protein or polypeptide part, said desired protein or polypeptide part then being cleaved off from the rest of the fusion protein or polpeptide either while the latter is adsorbed to the IgG-supporting carrier or after desorption thereof from the carrier. Also a hybrid vector for use herein, a method and an expression vector for its preparation and a host organism transformed by said hybrid vector are disclosed.
摘要翻译：通过构建重组载体来产生和选择性分离期望的蛋白质或多肽或其衍生物的方法，所述重组载体包含编码所述所需蛋白质或多肽的DNA序列，所述DNA序列可操作地连接于编码蛋白A的DNA序列或其活性多肽片段或任何其它能够结合免疫球蛋白恒定区的大分子，使得所述DNA序列一起编码所述所需蛋白质或多肽与所述蛋白A，其活性多肽片段或大分子之间的IgG结合融合产物; 用所述重组载体转化相容性宿主，使得编码所述融合蛋白或多肽的组合DNA序列可以由宿主表达，并在合适的生长培养基中培养转化的宿主以产生所述融合蛋白或多肽; 通过吸附到IgG载体材料上选择性分离所述融合蛋白或多肽; 并且任选地从所述IgG载体载体解吸所述融合蛋白或多肽，所述融合蛋白或由所述组合DNA序列编码的多肽任选地包含所述蛋白A部分和所述所需蛋白质或多肽部分之间的唯一切割位点，所述所需蛋白质或多肽部分然后从融合蛋白或多肽的其余部分切割，而后者被吸附到IgG载体载体上或者在其从载体解吸后。还公开了本文使用的杂交载体，用于其制备的方法和表达载体以及由所述杂交载体转化的宿主生物体。

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