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Renaturation and Purification of Biologically Active Recombinant Human Macrophage Colony-Stimulating Factor Expressed in E. Coli

Bio/Technology volume 7, pages 710–715 (1989)Cite this article

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.

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Author notes

  1. Robert Halenbeck: Corresponding author.

Authors and Affiliations

  1. Department of Protein Chemistry, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA, 94608

    Robert Halenbeck & Kirston Koths

  2. Department of Molecular Biology, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA, 94608

    Ernest Kawasaki

  3. Department of Immunology, Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA, 94608

    Joe Wrin

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  1. Robert Halenbeck
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  2. Ernest Kawasaki
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  3. Joe Wrin
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  4. Kirston Koths
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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

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