We have previously shown that the diphtheria toxin variant CRM197 (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.
Alessandra Stefan, Mattia Boiani, Luca Longanesi, Alejandro Hochkoeppler (2014). On-column refolding of diphtheria toxin variant CRM197 by different metal-chelating affinity chromatography matrices. JOURNAL OF CHEMISTRY AND CHEMICAL ENGINEERING, 8, 1135-1141 [10.17265/1934-7375].
On-column refolding of diphtheria toxin variant CRM197 by different metal-chelating affinity chromatography matrices
STEFAN, ALESSANDRA;HOCHKOEPPLER, ALEJANDRO
2014
Abstract
We have previously shown that the diphtheria toxin variant CRM197 (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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
