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Biotechnology and Bioprocess Engineering

, Volume 23, Issue 6, pp 670–678 | Cite as

An Efficient and Rapid Two-step Purification Method for Active Human Macrophage Colony-stimulating Factor from Escherichia coli

  • Jisu Lee
  • Jeongmin Lee
  • Eunha Hwang
  • Hye-Jung Kim
  • Sheunghun Lee
  • Hyerim LeeEmail author
  • Seong Yun Hwang
  • Sung-Jin Cho
  • Sun-Ju Yi
  • Kyunghwan Kim
Research Paper
  • 7 Downloads

Abstract

Macrophage colony-stimulating factor (M-CSF) is a hematopoietic growth factor that stimulates the proliferation and differentiation of mononuclear phagocytes. To be biologically active, M-CSF must form a homodimer linked by disulfide bridges. However, the refolding step of recombinant M-CSF is not only extremely laborious but also rate-limiting. Here, we describe an efficient and simplified method for refolding and purifying M-CSF from Escherichia coli. A truncated M-CSF (amino acids 36–181) was tagged with histidine, expressed, and then purified using nickel affinity columns under denaturing conditions. Our redox refolding buffer containing a mixture of reduced and oxidized glutathione and arginine correctly refolded M-CSF. Chromatography on Q-sepharose columns selectively purified the M-CSF dimer from the monomeric form. The dimeric nature of the purified M-CSF was confirmed using multi angle light scattering combined with high performance liquid chromatography. Moreover, cell proliferation and osteoclast differentiation assays using bone marrow-derived macrophages demonstrated that the recombinant M-CSF was biologically active ex vivo.

Keywords

M-CSF refolding osteoclastogenesis purification 

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Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jisu Lee
    • 1
  • Jeongmin Lee
    • 1
  • Eunha Hwang
    • 2
  • Hye-Jung Kim
    • 3
  • Sheunghun Lee
    • 1
  • Hyerim Lee
    • 1
    Email author
  • Seong Yun Hwang
    • 1
  • Sung-Jin Cho
    • 1
  • Sun-Ju Yi
    • 1
  • Kyunghwan Kim
    • 1
  1. 1.School of Biological Sciences, College of Natural SciencesChungbuk National UniversityCheongjuKorea
  2. 2.Korea Basic Science InstituteCheongjuKorea
  3. 3.New Drug Development CenterKBIO Osong Medical Innovation FoundationCheongjuKorea

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