Klyushnichenko V E, Tikhonov R V, Pechenov S E, Yakimov S A, Shingarova L N, Korobko V G, Wulfson A N
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklikho-Maklaya 16/10, Moscow, GSP-7, 117871, Russia.
Protein Expr Purif. 1998 Nov;14(2):261-6. doi: 10.1006/prep.1998.0952.
Two schemes for efficient and productive isolation for mutant human recombinant tumor necrosis factor-alpha (TNF-alpha R32H) from Escherichia coli cells were developed. The methods include membrane filtration, ion-exchange chromatography and gel filtration, and centrifugation with subsequent free-flow electrophoresis as an alternative procedure. The target product was obtained as active trimer with total yield more than 50% and greater than 98% purity according to PAGE, size-exclusion chromatography, HPLC, and HPCE.
开发了两种从大肠杆菌细胞中高效生产性分离突变型人重组肿瘤坏死因子-α（TNF-α R32H）的方案。这些方法包括膜过滤、离子交换色谱和凝胶过滤，以及离心后进行自由流电泳作为替代程序。根据聚丙烯酰胺凝胶电泳（PAGE）、尺寸排阻色谱、高效液相色谱（HPLC）和高效毛细管电泳（HPCE），目标产物以活性三聚体形式获得，总收率超过50%，纯度大于98%。
