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KMID : 1094720160210030446
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Biotechnology and Bioprocess Engineering
2016 Volume.21 No. 3 p.446 ~ p.452
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Cloning and expression of recombinant macrophage-colony stimulating factor - A progressive strategy for economical production
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Sohn Hong-Moon
Ko Young-Jong
Park Min-Eon
Kim Bo-Ra
Kim Ok-Joon
Kim Dong-Hwi
Moon Young-Lae
Lim Won-Bong
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Abstract
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Macrophage-colony stimulating factor (M-CSF) has been reported to be required for the proliferation and differentiation of macrophages from hematopoietic progenitor cells. Recently, recombinant M-CSF (rM-CSF) became widely used as a biological research reagent in bone marrow stimulations, vaccine development, gene therapy approaches, and stem cell mobilization. rM-CSF is a glycoprotein that activates and enhances the differentiation and survival of macrophages, which play a key role in the osteoclastogenetic response. Here, we describe the construction of the gene encoding rM-CSF, its cloning, and expression in Escherichia coli, as well as the purification of rM-CSF protein, and its activity in a biological assay in mouse bone marrow cells. Our results show that the combination of experimental strategies employed to obtain recombinant rM-CSF can yield a biologically active protein, and may be useful when scaling-up production of other biologically similar proteins.
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KEYWORD
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M-CSF, osteoclastogenesis, macrophage, cloning
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