Process development for production of recombinant human insulin-like growth factor-I in Escherichia coli

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1038/sj.jim.2900773
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FormatText
TypeArticle
Journal titleJournal of Industrial Microbiology and Biotechnology
ISSN1367-5435
1476-5535
Volume24
Issue2
Pages94–99
Abstract
Fed-batch cultures were carried out to overproduce human insulin-like growth factor I (IGF-I) in Escherichia coli. The effects of carbon sources (glucose or glycerol) and induction time on cell growth and IGF-I production were investigated in more detail. Glycerol was a better carbon source than glucose for IGF-I production in fed-batch culture. Induction at the mid-exponential phase with glycerol as a carbon source in the pH-stat fed-batch culture was optimal for IGF-I production. Under this condition, 2.8 g L[SUP-1] of fusion IGF-I was produced as inclusion bodies. We have also developed downstream processing for preparative scale purification of IGF-I from the fusion protein produced by the fed-batch culture using glycerol as a carbon source. After the fusion protein expressed was solubilized in 8 M urea and cleaved with hydroxylamine, the released IGF-I was purified by cation exchange chromatography, refolding and preparative scale reverse phase HPLC (rp-HPLC) to give recombinant IGF-I of >98% purity. The biological activities of the purified IGF-I were measured and found to be identical to those of commercial IGF-I.
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LanguageEnglish
Peer reviewedYes
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NPARC number23001828
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Record identifier983f89c3-6b7b-46b7-a7f0-6556540cf123
Record created2017-04-13
Record modified2017-04-13
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