Expression and Purification of Tag-removed Human IL37 by Digestion on Beads in Escherichia coli

Title:Expression and Purification of Tag-removed Human IL37 by Digestion on Beads in Escherichia coli

Volume: 25 Issue: 11

Author(s): Daichao Wu, Guoqing Li, Yao Ma, Jin Liu, Yukun Li, Juan Wang, Junhui Bai, Sijie Tan*, Meixiang Li*Yongheng Chen*

Affiliation:

• Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001,China

• Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001,China

• Laboratory of Structural Biology, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, XiangYa Hospital, Central South University, Changsha, Hunan 410008,China

Keywords: Soluble expression, purification, interleukin 37, Escherichia coli, digestion on beads, inflammatory diseases.

Abstract: Background: Human Interleukin 37 (IL37), a unique anti-inflammatory cytokine of IL1 family member, plays critical roles in innate and adaptive immunity and inflammation.

Objective: Preparation of high purity and tag-removed recombinant IL37 protein (rIL37) is critical for its clinical application.

Method: In this study, we constructed an N-terminal cleavable GST-fused IL37 expression vector for recombinant expression.

Results: Subsequent to transformation and optimization of the induction temperature, the soluble expression level of rIL37 was 306.5 mg/L of culture medium at 18 °C induction in Escherichia coli. Meanwhile, rIL37 was digested on beads by GST-HRV3C protease during GST affinity chromatography. After further purification, the purity of rIL37 was higher than 99 %. Finally, the antiinflammatory activity of tag-removed protein was verified by the results showing that rIL37 suppressed IL1β production in PBMCs.

Conclusion: This work presents a protocol to produce high purity and tag-removed rIL37 with antiinflammatory activity, which provides the firm basis for advancing clinical application in human IL37-related inflammatory diseases.

Cite this article as:

Wu Daichao , Li Guoqing , Ma Yao , Liu Jin , Li Yukun , Wang Juan , Bai Junhui , Tan Sijie *, Li Meixiang *, Chen Yongheng *, Expression and Purification of Tag-removed Human IL37 by Digestion on Beads in Escherichia coli, Protein & Peptide Letters 2018; 25 (11) . https://dx.doi.org/10.2174/0929866526666181128124028