The Biomedical Research Centre, University of British Columbia, Vancouver, British Columbia, Canada.
PLoS One. 2012;7(11):e49891. doi: 10.1371/journal.pone.0049891. Epub 2012 Nov 14.
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.
人粒细胞巨噬细胞集落刺激因子（hGM-CSF）是一种造血生长因子和前炎症细胞因子。重组 hGM-CSF 不仅是一种研究工具，而且是一种生物治疗药物。然而，在大肠杆菌中表达的 rhGM-CSF 已知会形成错误折叠、聚集的蛋白质包涵体。从包涵体中复性和随后纯化 rhGM-CSF 是困难的，因为产生的生物活性蛋白产量低。在这里，我们描述了一种从包涵体中分离、复性和纯化生物活性 rhGM-CSF 的方法。该方法简单易行，不需要在蛋白质复性或纯化方面有丰富的经验，并且使用标准的实验室设备。
