Abstract
Interleukin-18 binding protein (IL-18BP) is a natural IL-18 inhibitor in vivo, which can effectively neutralize IL-18 and inhibit the inflammatory signaling pathway induced by IL-18, thus playing an anti-inflammatory role. Traditional production methods primarily rely on eukaryotic animal cell expression systems, which often entail complex processes, lower yields, and increase production costs. In this study, we present a novel approach for expressing IL-18BP fusion protein using the Escherichia coli (E. coli) system. The N-terminal segment of IL-18BP was fused with the small ubiquitin-related modifier (SUMO) tag, enabling soluble expression, while the C-terminal segment was fused with the human IgG1 Fc fragment to prolong its in vivo lifespan. Through screening, we obtained a high-expression engineering strain from a single colony and developed optimized protocols for fermentation and purification of the recombinant SUMO-IL-18BP-Fc protein. The SUMO tag was subsequently cleaved using SUMO protease, and the purified recombinant human IL-18BP-Fc (rhIL-18BP-Fc) exhibited a purity exceeding 90% with a yield of 1 g per liter of bacterial solution. The biological activities and underlying mechanisms of rhIL-18BP-Fc were evaluated using cell lines and a mouse model. Our results demonstrated that rhIL-18BP-Fc effectively inhibited IL-18-stimulated IFN-γ production in KG-1a cells in vitro and ameliorated DSS-induced ulcerative colitis in mice. In conclusion, we successfully employed the SUMO fusion system to achieve high-level production, soluble expression, and prolonged activity of rhIL-18BP-Fc in E. coli. These findings lay the groundwork for future large-scale industrial production and pharmaceutical development of rhIL-18BP-Fc protein.
Key points
• Effective expression, fermentation, and purification of bioactive rhIL-18BP-Fc protein in E. coli.
• The rhIL-18BP-Fc protein has a great potential for the therapy of ulcerative colitis by inhibiting the expression of inflammatory factors.
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All data generated or analyzed during this study are included in this published article (and its supplementary information files).
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Funding
This work was supported by the Guangdong Provincial Department of Education (2021ZDZX2034) and the Guangdong Basic and Applied Basic Research Foundation (2022A1515010028).
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L.G. and X.C. performed experiments, analyzed figures, and wrote the manuscript. N.T. provided materials. H.Z. and W.L. performed partial experiments. J.Z. and Y.X. conceived the project, supervised experiments, analyzed data, and wrote the manuscript. All authors read and approved the final manuscript.
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Guo, L., Chen, X., Zeng, H. et al. Production of recombinant human long-acting IL-18 binding protein: inhibitory effect on ulcerative colitis in mice. Appl Microbiol Biotechnol 107, 7135–7150 (2023). https://doi.org/10.1007/s00253-023-12806-8
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DOI: https://doi.org/10.1007/s00253-023-12806-8