Process for preparing and purifying alpha-interferon
    4.
    发明授权
    Process for preparing and purifying alpha-interferon 失效
    制备和纯化α-干扰素的方法

    公开(公告)号:US5196323A

    公开(公告)日:1993-03-23

    申请号:US713618

    申请日:1991-06-12

    IPC分类号: A61K38/00 C07K14/56

    CPC分类号: C07K14/56 A61K38/00

    摘要: A process for the preparation and purification of recombinant Interferon-.alpha. is disclosed. The invention is directed to a process comprising the following steps: cultivating E. coli containing the interferon gene for a growth period during which not more than 5% methionine-interferon is formed; extracting and concentrating the expressed interferon; subjecting the preliminarily purified material to Tandem Chromatography, wherein the Tandem Chromatography comprises separation on a cellulose column followed by an anti-alpha-interferon monoclonal antibody affinity column; subjecting the thus purified material to isoelectric precipitation of impurities at about pH 4.0 to about pH 4.8; and purifying the interferon by chromatography on a high performance cation exchange column using a volatile buffer, wherein the purified interferon is non-immunogenic when administered parenterally to a human.

    摘要翻译: 公开了重组干扰素-α的制备和纯化方法。 本发明涉及一种包括以下步骤的方法:培养含有干扰素基因的大肠杆菌生长期,其中不超过5%的甲硫氨酸 - 干扰素形成; 提取和浓缩表达的干扰素; 将预先纯化的材料进行串联层析,其中串联色谱法包括在纤维素柱上分离,随后是抗α-干扰素单克隆抗体亲和柱; 使如此纯化的材料经受大约pH 4.0至约pH 4.8的杂质的等电沉淀; 并使用挥发性缓冲液在高性能阳离子交换柱上通过色谱法纯化干扰素,其中当肠胃外给予人时,纯化的干扰素是非免疫原性的。