Renaturation and Purification of Biologically Active Recombinant Human Macrophage Colony-Stimulating
Author: ["Robert Halenbeck","Ernest Kawasaki","Joe Wrin","Kirston Koths"]
Publication: Bio/Technology
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Abstract
Recombinant human macrophage colony-stimulating factor (M-CSF), a disulfide-linked dimeric protein containing 18 cysteines, has been purified in monomeric form from E. coli and renatured to generate fully active dimers with an overall yield of 25 percent. M-CSF monomers were quantitatively refolded at concentrations as high as 0.7 mg/ml. Residual contaminants, including endotoxin, were removed from the refolded M-CSF by hydrophobic interaction chromatography. The kinetics of refolding and the characteristics of the renatured product were determined by measuring (1) molecular size, (2) biological activity, and (3) reactivity with a novel neutralizing monoclonal antibody specific for the dimeric form of M-CSF. The results show that refolded M-CSF from E. coli closely resembles both native and recombinant M-CSF produced by mammalian cells.
Cite this article
Halenbeck, R., Kawasaki, E., Wrin, J. et al. Renaturation and Purification of Biologically Active Recombinant Human Macrophage Colony-Stimulating Factor Expressed in E. Coli. Nat Biotechnol 7, 710–715 (1989). https://doi.org/10.1038/nbt0789-710
