Scale-up process for expression and renaturation of recombinant human epidermal growth factor from Escherichia coli inclusion bodies.
- Title
- Scale-up process for expression and renaturation of recombinant human epidermal growth factor from Escherichia coli inclusion bodies.
- Author
- 정일엽
- Keywords
- L-cystine,; fed-batch; recombinant EGF; refolding
- Issue Date
- 2000-06
- Publisher
- Portland Press Ltd
- Citation
- BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, v. 31, no. 3, page. 245-248
- 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.
- URI
- https://iubmb.onlinelibrary.wiley.com/doi/full/10.1042/BA19990101https://repository.hanyang.ac.kr/handle/20.500.11754/161677
- DOI
- 10.1042/BA19990101
- Appears in Collections:
- COLLEGE OF SCIENCE AND CONVERGENCE TECHNOLOGY[E](과학기술융합대학) > MOLECULAR AND LIFE SCIENCE(분자생명과학과) > Articles
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