目的：制备符合临床要求的高纯度新型人肿瘤坏死因子（ｎｈＴＮＦ），建立简单、高效的纯化途径．方法：用本室构建的ｎｈＴＮＦ基因工程菌，经发酵、收菌、裂菌、硫酸铵盐析，Ｑ－Ｓｅｐｈａｒｏｓｅ及Ｓ－Ｓｅｐｈａｒｏｓｅ层析，得到高活性、高纯度的ｎｈＴＮＦ单一色谱峰，比活性达１×１０９Ｕ／ｍｇ蛋白．结果：ｎｈＴＮＦ纯度达９８％以上，活性回收率为７９％．结论：纯化方法简单，且达到较好效果，为ｎｈＴＮＦ的大规模制备和临床应用打下基础． OBJECTIVE: To prepare novel high purity human tumor necrosis factor (nhTNF) which meets clinical requirements and to establish a simple and efficient purification method. Methods: The nhTNF genetically engineered bacteria constructed by our laboratory were used to obtain the single peak of nhTNF with high activity and purity by fermentation, bacteria collection, fission, ammonium sulfate salting-out, Q-Sepharose and S-Sepharose chromatography. Up to 1 × 109U / mg protein. Results: The purity of nhTNF was above 98% and the activity recovery was 79%. Conclusion: The purification method is simple and achieves good results, laying a foundation for the large-scale preparation and clinical application of nhTNF.