On-column refolding of recombinant human interferon-gamma inclusion bodies by expanded bed adsorption chromatography.
A refolding strategy was described for on-column refolding of recombinant human interferon-gamma (rhIFN-gamma) inclusion bodies by expanded bed adsorption (EBA) chromatography. After the denatured rhIFN-gamma 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-gamma protein was recovered by the elution in packed bed mode. It was demonstrated that the denatured rhIFN-gamma protein could be efficiently refolded by this method with high yield. Under appropriate experimental conditions, the protein yield and specific activity of rhIFN-gamma was up to 52.7% and 8.18 x 10(6) IU/mg, respectively.
Jin
Ting
T
Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China.
Guan
Yi-Xin
YX
Yao
Shan-Jing
SJ
Lin
Dong-Qiang
DQ
Cho
Man-Gi
MG
D000327:Adsorption; D019149:Bioreactors; D002411:Cation Exchange Resins; D002845:Chromatography / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry*; D007371:Interferon-gamma / Q000737:chemistry*; D017510:Protein Folding; D011994:Recombinant Proteins
D002411:Cation Exchange Resins; D011994:Recombinant Proteins; D007371:Interferon-gamma