Affinity-assisted in vivo folding of a secreted human peptide hormone in Escherichia coli

Nature Biotechnology
E SamuelssonM Uhlén

Abstract

We show that coexpression of a specific binding protein in Escherichia coli can significantly improve the relative yields of correctly folded human insulin-like growth factor I (IGF-I). A glutathione redox buffer was used during growth to allow formation and breakage of disulfide bonds in the periplasm of the bacterial host. Both the binding protein and the peptide hormone were produced as affinity fusions, which allowed purification of the in vivo formed heterodimer by alternative affinity purification methods. The use of affinity-assisted in vivo folding has general implications for expression, folding, and purification of recombinant proteins.

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Citations

Apr 6, 2004·Journal of Biotechnology·F J M MergulhãoG A Monteiro
Mar 15, 2005·Biotechnology Advances·F J M MergulhãoG A Monteiro
Nov 5, 1997·Protein Expression and Purification·J NilssonP A Nygren

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