重组人突变型TNF-α在大肠杆菌中的诱导表达及高效纯化

Expression and Purification of a Recombinant Human Mutated TNF-α

建立重组人突变型ＴＮＦ－α的纯化工艺，获得ＴＮＦ－α纯品，为下游规模生产打基础。方法：诱导表达重组 ＴＮＦ－α突变体蛋白，用 ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ ＦＦ、ＣＭ－Ｓｅｐｈａｒｏｓｅ ＦＦ两步层析结合小范围线性梯度洗脱法纯化。结果：所得重组蛋白的纯度达９８％以上，比活达１．７×１０９ｕ／ｍｇ蛋白，无热原。结论：此纯化方法可对重组ＴＮＦ－α突变体蛋白进行高效快速纯化，易于实现规模化生产。

Purpose: The aim was to establish a procedure for the purification of a recombinant human mutated TNF-α- [Lys2] hTNF- α. Methods: The [Lys2] hTNF-α was purified with DEAE-Sepharose FF and CMSepharose FF iron exchange resins, eluted with linar gradient concentration (0-100 mmol/L) of NaCl. Results: The purity of the [Lys2]hTNF-α was more than 98%, and bioactivity was 1. 7 × 109 u/mg. Conclusion: The procedure established is an effective and fast method that is suitable for the purification of [Lys2]hTNF-α on a large scale.