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

[ Vol. 25 , Issue. 11 ]

Author(s):

Daichao Wu, Guoqing Li, Yao Ma, Jin Liu, Yukun Li, Juan Wang, Junhui Bai, Sijie Tan*, Meixiang Li* and Yongheng Chen*   Pages 996 - 1002 ( 7 )

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.

Keywords:

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

Affiliation:

Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study, University of South China, Hengyang, Hunan 421001, Network Information Center, University of South China, Hengyang, Hunan 421001, Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Department of Histology and Embryology, Institute of Clinical Anatomy & Reproductive Medicine, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, Laboratory of Structural Biology, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, XiangYa Hospital, Central South University, Changsha, Hunan 410008

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