title: Purification of soluble human epidermal growth factor (hEGF) from recombinant Escherichia coli culture broth by using expanded-bed adsorption chromatography.
abstract: Human epidermal growth factor (hEGF) secreted by recombinant Escherichia coli was purified from culture broth by expanded-bed adsorption (EBA) chromatography, strong anion-exchange chromatography and finally preparative reversed-phase HPLC (RP-HPLC). The EBA chromatography step simultaneously captured the hEGF by cationic exchanger and removed the cellular biomass from the diluted culture broth. This step was carried out at high throughput, and resulted in a high yield (>90%) and a purification factor of approx. 20-fold to >80% purity. Its process performance was well maintained during a 16-fold scale-up. After the successive purification steps of anion-exchange chromatography and RP-HPLC, the overall yield was approx. 84% and the purity was satisfactory (>99.5%). It was concluded that the purification process was very efficient and scaleable, warranting its implementation in large-scale manufacturing.
authors.affiliation: Institute of Bioscience and Biotechnology, DaeWoong Pharmaceutical Corp., YongIn, South Korea.
authors.forename: Yoon Seok
authors.initials: YS
authors.lastname: Lee
authors.forename: Chang Woo
authors.initials: CW
authors.lastname: Suh
authors.forename: Seung Kook
authors.initials: SK
authors.lastname: Park
authors.forename: Eun Kyu
authors.initials: EK
authors.lastname: Lee
mesh_terms: D000327:Adsorption; D002845:Chromatography / Q000379:methods*; D002847:Chromatography, Agarose; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D004815:Epidermal Growth Factor / Q000096:biosynthesis / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D006801:Humans; D011994:Recombinant Proteins / Q000302:isolation & purification; D012995:Solubility
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor