Dorin G, Thomson J, Hanisch W
Cetus Corporation, Emeryville, California 94608.
Biotechnol Prog. 1990 Nov-Dec;6(6):494-7. doi: 10.1021/bp00006a013.
Recombinant tumor necrosis factor (TNF) is expressed in Escherichia coli as a soluble intracellular protein. A purification process is described that incorporates a hydrophilic membrane (cellulosic) separation followed by a hydrophobic one (PTFE). The hydrophilic step is a traditional one in that species are separated primarily on the basis of size. The hydrophobic step separates species on the basis of parameters apparently not related to size. By combining these two steps, an increase in TNF purity of 7-10-fold can be achieved with a yield of 50%. The effects of cellular debris and pH on selectivity and recovery of the hydrophobic filtration step are discussed.
重组肿瘤坏死因子（TNF）在大肠杆菌中作为可溶性细胞内蛋白表达。本文描述了一种纯化方法，该方法包括先进行亲水膜（纤维素）分离，然后进行疏水膜（聚四氟乙烯）分离。亲水步骤是传统的分离步骤，主要根据大小对不同物质进行分离。疏水步骤则根据明显与大小无关的参数对不同物质进行分离。通过将这两个步骤相结合，TNF的纯度可提高7至10倍，产率为50%。文中还讨论了细胞碎片和pH值对疏水过滤步骤的选择性和回收率的影响。
