title: [Refolding and purification of recombinant human granulocyte colony-stimulating factor from Escherichia coli by using protein folding liquid chromatography].
abstract: Recombinant human granulocyte colony-stimulating factor (rhG-CSF) in the form of inclusion bodies expressed in Escherichia coli (E. coli) was simultaneously refolded and purified using protein folding liquid chromatography (PFLC). Cu2+ -iminodiacetic acid (IDA) Sepharose was selected as the stationary phase for immobilized metal ion affinity chromatography. rhG-CSF was purified and the aggregates were diminished under a linear gradient elution of imidazole in the presence of a suitable concentration of urea. Using only one PFLC run, the refolded rhG-CSF had a specific bioactivity of 1.8 x 10(8) IU/mg and a purity of 97%, with the mass recovery of 32%.
authors.affiliation: Key Laboratory of Separation Science in Shaanxi Province, Institute of Modern Separation Science, Department of Chemistry, Northwest University, Xi' an 710069, China. czwang@nwu.edu.cn
authors.forename: Chaozhan
authors.initials: C
authors.lastname: Wang
authors.forename: Lili
authors.initials: L
authors.lastname: Wang
authors.forename: Xindu
authors.initials: X
authors.lastname: Geng
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D003300:Copper / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism / Q000652:urine; D006801:Humans; D007098:Imino Acids / Q000737:chemistry; D017510:Protein Folding; D011994:Recombinant Proteins
chemical_list: D007098:Imino Acids; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D003300:Copper; C008109:iminodiacetic acid