title: A simplified method for the efficient refolding and purification of recombinant human GM-CSF.
abstract: Human granulocyte macrophage colony-stimulating factor (hGM-CSF) is a haematopoietic growth factor and proinflammatory cytokine. Recombinant hGM-CSF is important not only as a research tool but also as a biotherapeutic. However, rhGM-CSF expressed in E. coli is known to form inclusion bodies of misfolded, aggregated protein. Refolding and subsequent purification of rhGM-CSF from inclusion bodies is difficult with low yields of bioactive protein being produced. Here we describe a method for the isolation, refolding and purification of bioactive rhGM-CSF from inclusion bodies. The method is straightforward, not requiring extensive experience in protein refolding nor purification and using standard laboratory equipment.
authors.affiliation: The Biomedical Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Christy A
authors.initials: CA
authors.lastname: Thomson
authors.forename: Melanie
authors.initials: M
authors.lastname: Olson
authors.forename: Linda M
authors.initials: LM
authors.lastname: Jackson
authors.forename: John W
authors.initials: JW
authors.lastname: Schrader
mesh_terms: D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000737:chemistry* / Q000302:isolation & purification*; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D013058:Mass Spectrometry; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor