Facilitated in vitro refolding of human recombinant insulin-like growth factor I using a solubilizing fusion partner.
We describe a new approach to refolding recombinant proteins in which an affinity fusion partner, consisting of two IgG-binding domains (ZZ) derived from staphylococcal protein A, is used to solubilize misfolded molecules before, during and after reduction and reoxidation. We show that human insulin-like growth factor I (IGF-I) can be refolded as a fusion protein at a concentration as high as 1-2 mg/ml without the use of denaturing agents. A process scheme suitable for large scale application is described in which the yield of correctly folded human IGF-I with full biological activity is substantially increased.
Department of Biochemistry, Royal Institute of Technology, Stockholm, Sweden.
E
E
Samuelsson
H
H
Wadensten
M
M
Hartmanis
T
T
Moks
M
M
Uhlén
D007074:Immunoglobulin G / Q000737:chemistry; D066298:In Vitro Techniques; D007334:Insulin-Like Growth Factor I / Q000737:chemistry*; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry
D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D007334:Insulin-Like Growth Factor I