[Refolding and purification of the huGM-CSF(9-127)-IL-6(29-184) fusion protein].
The huGM-CSF(9-127)-IL-6(29-184) fusion protein was precipitated on column when being purified by Q Sepharose H.P. ion exchange chromatography after renaturation by dilution. To solve this problem, a novel purification and refolding strategy was adopted. Inclusion bodies was first purified by Q Sepharose H.P. ion exchange in 8 mol/L urea, followed by in situ refolding on column by Sephacryl S-200. Renatured fusion protein was obtained in a purity of more than 95%. It was showed that the method of refolding on gel filtration column is efficient, with relative refolding rate at 80%. By the whole procedure, refolding and purification of recombinant protein can be performed within one day. This strategy is also promising to be applied in large scale purification and refolding of recombinant protein from inclusion bodies in E. coli.
Department of Molecular Biology, Institute of Medical Biology, CAMS and PUMC, Kunming 650118, China.
Qiang-Ming
QM
Sun
Hong-Yan
HY
Liu
Chang-Bai
CB
Dai
Yan-Bing
YB
Ma
Mao-Sheng
MS
Sun
Wei-Ming
WM
Xu
D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000737:chemistry / Q000302:isolation & purification*; D015850:Interleukin-6 / Q000737:chemistry / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*
D015850:Interleukin-6; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor