文献类型：期刊文献

英文题名：Refolding and purification of Apo2L/TRAIL produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli

作者：Shen, XL; Xia, XX; Zhang, Y; Liu, JW; Wei, DZ; Yang, SL

机构：[1]E China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2003

卷号：25

期号：24

起止页码：2097

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000187002700011)】；

语种：英文

外文关键词：high-cell-density cultivation; inclusion bodies; purification; refolding; tumor necrosis factor-related apoptosis-inducing ligand

摘要：As a new member of tumor necrosis factor (TNF) superfamily, TNF-related apoptosis- inducing ligand (Apo2L/TRAIL) was produced mainly as inclusion bodies by recombinant Escherichia coli with a temperature-inducible expression system. High concentrations of both biomass (65 g dry cells l(-1)) and inactive TRAIL (4.8 g l(-1)) were obtained by applying a high-cell-density cultivation procedure. After the inclusion bodies were washed and solubilized, TRAIL refolded when at 1 mg ml(-1) by a simple pulse dilution method with a 35% yield. Renatured TRAIL was purified to electrophoretic homogeneity by one-step immobilized metal affinity chromatography. The purified TRAIL showed strong cytotoxicity activity against human pancreatic 1990 tumor cells, with ED50 about 1.6 mug ml(-1).