Biotechnology Letters

, Volume 29, Issue 9, pp 1323–1327 | Cite as

Expression, purification and characterization of soluble human interleukin-6 receptor α-chain and its mutant protein in Escherichia coli

  • Jing Wang
  • Zhenhui Yang
  • Ming Shi
  • Yan Li
  • Ning Guo
  • Beifen Shen
  • Jiannan Feng
Original Research Paper
First Online:
Received:
Revised:
Accepted:
  • 157 Downloads

Abstract

To investigate the function of the N-terminal immunoglobulin (Ig)-like domain of the human interleukin-6 receptor α-chain (hIL-6R), we constructed a soluble human interleukin-6 receptor (shIL-6R) (named EC05, amino acids 20–354) and soluble variants of the shIL-6R lacking the Ig-like domain (named EC70, amino acids 105–354). The two extracellular portions of hIL-6R were expressed as soluble fusion proteins with thioredoxin in Escherichia coli and purified by using Ni-NTA agarose. Western blot showed that purified proteins were immunoreactive with the antibody against hIL-6R. They also possessed specific binding activity with human interleukin-6 (hIL-6) in ELISA analysis.

Keywords

Escherichia coli Human interleukin-6 receptor Immunoglobulin domain Protein purification Soluble fusion expression 
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Notes

Acknowledgements

The authors thank Prof. Hirano (Osaka University, Osaka, Japan) for supplying the full length of the extracellular hIL-6R plasmid. This study was supported by National Sciences Fund (No. 30490240), “973” Fund (2003CB515508) and Beijing National Sciences Fund (No. 5062037) of P. R. China.

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Jing Wang
    • 1
  • Zhenhui Yang
    • 1
  • Ming Shi
    • 1
  • Yan Li
    • 1
  • Ning Guo
    • 1
  • Beifen Shen
    • 1
  • Jiannan Feng
    • 1
  1. 1.Department of Molecular ImmunologyInstitute of Basic Medical SciencesBeijingP.R. China

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