On‐column refolding of recombinant human interferon‐γ inclusion bodies by expanded bed adsorption chromatography

A refolding strategy was described for on-column refolding of recombinant human interferon-γ (rhIFN-γ) inclusion bodies by expanded bed adsorption (EBA) chromatography. After the denatured rhIFN-γ protein bound onto the cation exchanger of STREAMLINE SP, the refolding process was performed in expanded bed by gradually decreasing the concentration of urea in the buffer and the refolded rhIFN-γ protein was recovered by the elution in packed bed mode. It was demonstrated that the denatured rhIFN-γ protein could be efficiently refolded by this method with high yield. Under appropriate experimental conditions, the protein yield and specific activity of rhIFN-γ was up to 52.7% and 8.18 × 106 IU/mg, respectively. © 2005 Wiley Periodicals, Inc.