Affinity-assisted in vivo folding of a secreted human peptide hormone in Escherichia coli.
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.
Samuelsson
E
E
Department of Biochemistry and Biotechnology, KTH, Stockholm, Sweden.
Jonasson
P
P
Viklund
F
F
Nilsson
B
B
Uhlén
M
M
D000345:Affinity Labels; D015153:Blotting, Western; D002021:Buffers; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D018970:Insulin-Like Growth Factor Binding Protein 1 / Q000235:genetics; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010084:Oxidation-Reduction; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D000345:Affinity Labels; D002021:Buffers; D018970:Insulin-Like Growth Factor Binding Protein 1; D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I