Soluble expression and purification of bioactive interleukin 33 in E. coli

Bich Hang Do, Sangsu Park, Grace G. Kwon, Minh Tan Nguyen, Hyo Jeong Kang, Jung A. Song, Jiwon Yoo, Anh Ngoc Nguyen, Jaepyeong Jang, Mihee Jang, Sunju Lee, Seoungjun So, Sungrak Sim, Jonghwa Jin, Kyung Jin Lee, Mark J. Osborn, Han Choe

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Interleukin-33 (IL-33) is one of the important alarmins of the immune system and possesses dual functions as an anti- or pro-inflammatory molecule. The production of this cytokine in E. coli is hampered by the insoluble expression in the cytoplasm, resulting in inclusion body formation. In this study, the expression of IL-33 was optimized by fusing the N-terminus of IL-33 with several solubilizing tags that act as chaperones for proper protein folding: maltose binding protein (MBP), b´a´ domain of protein disulfide isomerase (PDIb´a´) and glutathione Stransferase (GST). The expression of the fusion proteins was stimulated by 0.5 mM IPTG at different temperatures, 37, 30, 25, and 18°C. As a result, IL-33 was expressed highly and in soluble form in the cytoplasm of E. coli when fused with MBP or PDIb´a´ tags in the presence of 0.5 mM IPTG at 25 or 30°C. We describe a simple purification procedure of IL-33 from the PDIb´a´-IL-33 construct using immobilized metal affinity chromatography (IMACs) with supplementary of tobacco etch virus (TEV) protease for tag removal. The high bioactivity of purified IL-33 on the proliferation and activation of macrophages was confirmed by MTT and nitrite releasing assays using RAW 264.7 These data show an improved method for producing high grade and yield IL-33.

Original languageEnglish (US)
Pages (from-to)256-264
Number of pages9
JournalBiotechnology and Bioprocess Engineering
Volume22
Issue number3
DOIs
StatePublished - Jun 1 2017

Keywords

  • E. coli
  • human Interleukin 33 (hIL-33)
  • macrophage activation
  • macrophage proliferation
  • purification
  • soluble expression

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