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1. 发明授权

US06936437B2 Methods and compositions for production of recombinant peptides 失效
标题翻译：用于生产重组肽的方法和组合物
公开(公告)号：US06936437B2
公开(公告)日：2005-08-30
申请号：US10080919
申请日：2002-02-22
申请人： Lucia Irene Gonzalez-Villasenor
发明人： Lucia Irene Gonzalez-Villasenor
IPC分类号： C07K1/113 , C07K14/47 , C12P21/02 , C12P21/06 , C07K1/00 , C07K14/00 , C07K17/00 , C12P21/04
CPC分类号： C07K14/4716 , C07K1/113 , C12P21/02
摘要： This invention provides a method for solubilizing and recovering in bioactive form, a target polypeptide or protein that has been expressed from a host organism in insoluble form. The polypeptide is isolated from the host organism by standard procedures including disrupting the host cell to produce a lysate and recovering the precipitate from the lysate. The precipitate containing the polypeptide is solubilized in a non-denaturant and detergent free non-buffered solubilization solution containing sodium hydroxide between 8 and 10 mM, mannitol between 2 and 5 mM and lactose between 1 and 2 mM. The resultant solubilization preparation contains a biologically active target polypeptide between 1 and 4 mg per ml of solubilization solution. The solubilized polypeptide obtained in this particular manner constitutes the object of the invention. The bioactive solubilized polypeptide is then transferred into an appropriate buffered solution of the desired pH and purified following conventional chromatographic procedures.
摘要翻译：本发明提供一种以生物活性形式溶解和回收的方法，所述方法是从宿主生物以不溶形式表达的靶多肽或蛋白质。通过标准方法从宿主生物体分离多肽，包括破坏宿主细胞以产生裂解物并从裂解物中回收沉淀物。将含有多肽的沉淀溶解在含有8-10mM的氢氧化钠，2至5mM的甘露醇和1至2mM的乳糖的非变性和无去污剂的非缓冲溶解溶液中。所得的溶解制剂含有1至4mg / ml溶解溶液的生物活性目标多肽。以该特定方式获得的增溶多肽构成本发明的目的。然后将生物活性溶解的多肽转移到所需pH的适当缓冲溶液中，并按照常规色谱方法纯化。

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