title: Scale-up process for expression and renaturation of recombinant human epidermal growth factor from Escherichia coli inclusion bodies.
abstract: A cDNA encoding mature epidermal growth factor (EGF) was isolated and cloned into a pQE30 vector in which the His(6)-tagged EGF was expressed. pH-stat feeding of concentrated medium at the time of isopropyl beta-D-thiogalactoside induction and slug-feedings of the enriched medium during the induction resulted in a higher cell density and specific expression. Using a simple refolding protocol that consisted of 1 mM L-cysteine addition for a 1-h reduction followed by 5 mM L-cystine addition for oxidative refolding, we were able to convert nearly all EGF monomers into the oxidized form. Also, there folding aggregate was converted into the monomeric form. Approx. 50% overall yield was obtained from the dissolved inclusion bodies to a single peak under FPLC. We hope that the result of this study may provide information that is useful for the scale-up of the recombinant human EGF production process.
authors.affiliation: Department of Chemical Engineering, Hanyang University, Ansan, Korea 425-791.
authors.forename: J Y
authors.initials: JY
authors.lastname: Lee
authors.forename: C S
authors.initials: CS
authors.lastname: Yoon
authors.forename: I Y
authors.initials: IY
authors.lastname: Chung
authors.forename: Y S
authors.initials: YS
authors.lastname: Lee
authors.forename: E K
authors.initials: EK
authors.lastname: Lee
mesh_terms: D002455:Cell Division / Q000187:drug effects; D002460:Cell Line / Q000187:drug effects; D003470:Culture Media; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D002479:Inclusion Bodies / Q000235:genetics; D010084:Oxidation-Reduction; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor