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Biotechnology and Applied Biochemistry (2003) 38, (9–13) (Printed in Great Britain)

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Purification of soluble human epidermal growth factor (hEGF) from recombinant Escherichia coli culture broth by using expanded-bed adsorption chromatography

Yoon Seok Lee*, Chang Woo Suh†, Seung Kook Park* and Eun Kyu Lee†¹

*Institute of Bioscience and Biotechnology, DaeWoong Pharmaceutical Corp., YongIn, South Korea, and †Bioprocessing Research Laboratory, Department of Chemical Engineering, Hanyang University, 1271 Sa-dong, Ansan 425-791, South Korea

Key words: expanded-bed adsorption (EBA), Escherichia coli fermentation, human epidermal growth factor (hEGF), protein purification, scale-up.

Abbreviations used: hEGF, human epidermal growth factor; EBA, expanded-bed adsorption; RP-HPLC, reversed-phase HPLC; IPTG, isopropyl b-D-thiogalactoside.

¹To whom correspondence should be addressed (e-mail eklee@hanyang.ac.kr).

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.

Received 25 November 2002/3 February 2003; accepted 13 February 2003

Published as Immediate Publication 13 February 2003, DOI 10.1042/BA20020113

© 2003 Portland Press Ltd