产生复合人类长效IL-18结合蛋白:对小鼠性结肠炎的抑制作用
Lei Guo1,2, Xiuze Chen1, Haifeng Zeng3
1Department of Biotechnology, College of Basic Medical Science, Guangdong Medical University, Dongguan, 523808, China.
Applied microbiology and biotechnology
|September 28, 2023
概括
我们开发了一种新的大肠杆菌 (E. coli) 表达系统,用于产生复合的人类介质蛋白-18结合蛋白-Fc (rhIL-18BP-Fc). 这种方法确保了高产量和纯度,为性结肠炎治疗提供了潜力.
科学领域:
- 生物技术是生物技术.
- 再组合蛋白质的表达方式
- 炎症研究的研究.
背景情况:
- 介素-18结合蛋白 (IL-18BP) 是一种天然的IL-18抑制剂,对于调节炎症反应至关重要.
- 对于IL-18BP,传统的真核细胞表达系统是昂贵和低效的,限制了它的治疗潜力.
- 开发具有成本效益和可扩展的IL-18BP生产方法对于其临床应用至关重要.
研究的目的:
- 为人类IL-18BP与SUMO标签和人类IgG1 Fc片段融合在埃舍里希亚大肠杆菌 (E. coli) 中建立一种新的复合表达系统.
- 为了优化发酵和净化过程,以获得高产量,可溶性表达的rhIL-18BP-Fc融合蛋白.
- 在体外和体内评估纯化rhIL-18BP-Fc的生物活性和治疗潜力.
主要方法:
- 工程化大肠杆菌菌株,用于高水平的SUMO-IL-18BP-Fc融合蛋白的可溶性表达.
- 优化发酵和净化方案,以达到高纯度 (>90%) 和产量 (1 g/L).
- 使用SUMO蛋白酶进行SUMO标签切割,以获得生物活性rhIL-18BP-Fc.
- 在体外测试中使用KG-1a细胞来评估IL-18刺激的IFN-γ生产的抑制.
- 在体内研究使用DSS诱导的性结肠炎的小鼠模型.
主要成果:
- 在大肠杆菌中成功实现了rhIL-18BP-Fc的高水平可溶性表达和净化,纯度>90%和1g/L的产量.
- 在体外证明有效抑制IL-18诱导的IFN-γ在KG-1a细胞中的产生.
- 在小鼠模型中显示了DSS诱导的性结肠炎的显著改善,表明治疗疗效.
结论:
- 大肠杆菌中的SUMO融合系统能够高效地大规模生产生物活性的rhIL-18BP-Fc.
- rhIL-18BP-Fc表现出强大的抗炎活性,并对性结肠炎具有显著的治疗潜力.
- 这项研究为rhil-18BP-Fc.的工业生产和制药开发提供了基础.
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