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重组人肿瘤坏死因子α工程菌经发酵培养、破菌后，上清再经热处理、硫酸按分级沉淀，然后过SephadexG－25、DEANSepharoseCL-6B和Sephacry1S-100柱层析进行再纯化。结果显示，经多步层析纯化后rhTNFa回收率达27％，纯化倍数达236倍。连续纯化3批，纯度均达96％以上，比活性达3．9×106U／mg蛋白。提示所建立的纯化工艺稳定，为rhTNFα的基础研究和临床研究奠定了基础。
Recombinant human tumor necrosis factor alpha engineered bacteria were cultured and disrupted by bacteria, and the supernatant was heat-treated, precipitated by sulfuric acid, and then purified by column chromatography over Sephadex G-25, DEANSerose CL-6B and Sepaphary 1S-100. The results showed that the recovery of rhTNFa after multi-step chromatography was 27% and the purification factor was 236-fold. Purified continuously for 3 batches, the purity reached above 96%, and the specific activity reached 3.9×106 U/mg protein. It indicated that the established purification process was stable, which laid a foundation for basic research and clinical research of rhTNFα.