Preparative purification process for human furin
    25.
    发明授权
    Preparative purification process for human furin 有权
    人类弗林蛋白酶的制备纯化方法

    公开(公告)号:US08343748B2

    公开(公告)日:2013-01-01

    申请号:US12124390

    申请日:2008-05-21

    IPC分类号: C12N9/14 C07K1/18 C07K1/20

    摘要: Recombinant truncated human furin was expressed in CHO cells and concentrated approximately 50-fold by ultrafiltration and diafiltration. The concentrate was purified by column chromatography on CAPTO MMC™ (mixed cation exchange/hydrophobic interaction gel) resulting in a 30-50 fold purification factor and a yield of at least 60%. The at least 20% pure preparation obtained after CAPTO MMC™ (mixed cation exchange/hydrophobic interaction gel) chromatography had already a purification degree allowing on-column maturation of pro-VWF. Then an additional Arginine Sepharose chromatography purification was carried out. This two column process for purification of truncated human furin resulted in an almost pure furin preparation with a specific activity of approximately 290,000 U furin/mg protein and a yield of about 50%.

    摘要翻译: 重组截短的人弗林蛋白酶在CHO细胞中表达,并通过超滤和渗滤浓缩约50倍。 浓缩物通过CAPTO MMC TM(混合阳离子交换/疏水相互作用凝胶)上的柱色谱纯化,得到30-50倍纯化因子,产率至少为60%。 在CAPTO MMC TM(混合阳离子交换/疏水相互作用凝胶)色谱之后获得的至少20%的纯制剂已经具有允许亲VWF的柱上成熟的纯化度。 然后进行额外的精氨酸琼脂糖层析纯化。 用于纯化截短的人弗林蛋白酶的这种两柱方法产生几乎纯的弗林蛋白酶制剂,其具有约290,000U弗林蛋白/ mg蛋白质的比活性,约50%的产率。