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 μg ml−1.
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Shen, YL., Xia, XX., Zhang, Y. et al. Refolding and purification of Apo2L/TRAIL produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli . Biotechnology Letters 25, 2097–2101 (2003). https://doi.org/10.1023/B:BILE.0000007072.56577.0c
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DOI: https://doi.org/10.1023/B:BILE.0000007072.56577.0c