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A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF
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 misf...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3498172/ https://www.ncbi.nlm.nih.gov/pubmed/23166789 http://dx.doi.org/10.1371/journal.pone.0049891 |
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author | Thomson, Christy A. Olson, Melanie Jackson, Linda M. Schrader, John W. |
author_facet | Thomson, Christy A. Olson, Melanie Jackson, Linda M. Schrader, John W. |
author_sort | Thomson, Christy A. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3498172 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-34981722012-11-19 A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF Thomson, Christy A. Olson, Melanie Jackson, Linda M. Schrader, John W. PLoS One Research Article 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. Public Library of Science 2012-11-14 /pmc/articles/PMC3498172/ /pubmed/23166789 http://dx.doi.org/10.1371/journal.pone.0049891 Text en © 2012 Thomson et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Thomson, Christy A. Olson, Melanie Jackson, Linda M. Schrader, John W. A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF |
title | A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF |
title_full | A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF |
title_fullStr | A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF |
title_full_unstemmed | A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF |
title_short | A Simplified Method for the Efficient Refolding and Purification of Recombinant Human GM-CSF |
title_sort | simplified method for the efficient refolding and purification of recombinant human gm-csf |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3498172/ https://www.ncbi.nlm.nih.gov/pubmed/23166789 http://dx.doi.org/10.1371/journal.pone.0049891 |
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