title: Expression and purification of recombinant human granulocyte colony-stimulating factor in fed-batch culture of Escherichia coli.
abstract: Granulocyte colony-stimulating factor (G-CSF) is a cytokine that has multiple roles in hematopoietic cells such as the regulation of proliferation and differentiation. Here, we describe fed-batch culture, refolding, and purification of rhG-CSF. The suitability of urea or sarcosine for solubilizing inclusion bodies (IBs) was tested. It was observed that urea is more efficient for solubilizing and refolding IBs than sarcosine is. The purity of rhG-CSF and the removal percentage of the rhG-CSF isoforms during purification were increased by pH 5.5 precipitation. The purity and the yield of purified rhG-CSF were 99% and 0.5 g of protein per liter culture broth, respectively. Our protocols of recombinant protein purification using ion exchange chromatography and semipreparative high performance liquid chromatography of pH-precipitated refolded solution may be informative to the industrial scale production of biopharmaceuticals.
authors.affiliation: Department of Animal Biotechnology/Animal Resources Research Center, College of Animal Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin gu, Seoul, 143-701, South Korea.
authors.forename: Chang-Kyu
authors.initials: CK
authors.lastname: Kim
authors.forename: Jun-Ha
authors.initials: JH
authors.lastname: Choi
authors.forename: Seung-Bae
authors.initials: SB
authors.lastname: Lee
authors.forename: Sang-Mahn
authors.initials: SM
authors.lastname: Lee
authors.forename: Jae-Wook
authors.initials: JW
authors.lastname: Oh
mesh_terms: D061249:Batch Cell Culture Techniques; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D002479:Inclusion Bodies / Q000737:chemistry; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012521:Sarcosine / Q000737:chemistry; D014508:Urea / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D014508:Urea; D012521:Sarcosine