We have previously shown that the diphtheria toxin variant CRM 197 (cross-reacting material 197) can be overexpressed in Escherichia coli at high levels, yielding insoluble aggregates, which were solubilized using u...We have previously shown that the diphtheria toxin variant CRM 197 (cross-reacting material 197) can be overexpressed in Escherichia coli at high levels, yielding insoluble aggregates, which were solubilized using urea. This study reports a comparison of three matrices suitable for the purification and refolding of recombinant CRM197 by metal-chelating affinity chromatography, Moreover, we show that refolded CRM197 features enzymatic activity.展开更多
文摘We have previously shown that the diphtheria toxin variant CRM 197 (cross-reacting material 197) can be overexpressed in Escherichia coli at high levels, yielding insoluble aggregates, which were solubilized using urea. This study reports a comparison of three matrices suitable for the purification and refolding of recombinant CRM197 by metal-chelating affinity chromatography, Moreover, we show that refolded CRM197 features enzymatic activity.