Refolding and purification of Apo2L/TRAIL produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli |
| |
| Authors: | Ya-Ling Shen Xiao-Xia Xia Yue Zhang Jian-Wen Liu Dong-Zhi Wei Sheng-Li Yang |
| |
| Institution: | State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, PR China. |
| |
| Abstract: | 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 microg ml(-1). |
| |
| Keywords: | high-cell-density cultivation inclusion bodies purification refolding tumor necrosis factor-related apoptosis-inducing ligand |
| 本文献已被 PubMed SpringerLink 等数据库收录！ |
