Refolding and purification of the huGM-CSF(9-127)-IL-6(29-184) fusion protein.
- Author:
Qiang-Ming SUN
1
;
Hong-Yan LIU
;
Chang-Bai DAI
;
Yan-Bing MA
;
Mao-Sheng SUN
;
Wei-Ming XU
Author Information
1. Department of Molecular Biology, Institute of Medical Biology, CAMS and PUMC, Kunming 650118, China.
- Publication Type:Journal Article
- MeSH:
Granulocyte-Macrophage Colony-Stimulating Factor;
chemistry;
isolation & purification;
Interleukin-6;
chemistry;
isolation & purification;
Peptide Fragments;
chemistry;
isolation & purification;
Protein Folding;
Recombinant Fusion Proteins;
chemistry;
isolation & purification
- From:
Chinese Journal of Biotechnology
2002;18(3):291-294
- CountryChina
- Language:Chinese
-
Abstract:
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.
