Nanobody-Based Bispecific Neutralizer for Shiga Toxin-Producing E. coli
Nanobody-Based Bispecific Neutralizer for Shiga Toxin-Producing E. coli
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基于纳米抗体的双特异性中和剂，用于产志贺毒素大肠杆菌
DOI：
10.1021/acsinfecdis.1c00456
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发表时间：
2022
影响因子：
5.3
通讯作者：
Zhimeng Wu
中科院分区：
文献类型：
--
作者：
Zhongkai Lu;Zhicheng Liu;Xia Li;Xinfang Qin;Haofei Hong;Zhifang Zhou;Rol;J.Pieters;Jie Shi;Zhimeng Wu
目前，尚无针对食源性产志贺毒素大肠杆菌 (STEC) 感染的特效疗法，这种感染会导致严重胃肠炎和危及生命的溶血性尿毒症综合征 (HUS) 并发症。由于 STEC 与肠上皮的附着可能会增加宿主对志贺毒素的吸收和疾病的严重程度，因此我们受到启发，开发了一种能够同时阻断其志贺毒素和粘附素 intimin 的双特异性中和剂。针对志贺毒素 2 (Stx2B) 的 B 亚基和 Intimin (IntC280) 的 C 末端的两个纳米抗体通过基因融合在一起作为双特异性中和剂，并且可以在传统的大肠杆菌表达系统中高效生产。我们证明，双特异性形式的每个纳米抗体模块都显示出增强的抗原结合能力，并且即使在两种毒力因子同时存在的情况下，也能够在功能上中和 Stx2B 或 IntC280 与各自宿主受体的结合。此外，双特异性中和剂对于恶劣的储存条件和胃肠道极端 pH 值相对稳定。考虑到其简单、经济的生产和卓越的药物特性，我们相信基于纳米抗体的双特异性中和剂将更有利和更实用，可在发展中国家开发为对抗 STEC 的治疗方法。
Currently, no speci fi c therapeutics are available for.foodborne Shiga toxin-producing Escherichia coli (STEC).infections that cause severe gastroenteritis and life-threatening.complications of hemolytic uremic syndrome (HUS). As STEC.attachment to intestinal epithelium might increase the host.absorption of Shiga toxins and severity of the disease, we were.inspired to develop a bispeci fi c neutralizer capable of blocking its.Shiga toxin and adhesin intimin simultaneously. Two nanobodies.against the B subunit of Shiga toxin 2 (Stx2B) and the C terminus.of Intimin (IntC280) were genetically fused together as the.bispeci fi c neutralizer, and it can be e ffi ciently produced in a conventional E. coli expression system. We demonstrated that each of the.nanobody modules in the bispeci fi c format showed increased antigen binding capability and was able to functionally neutralize the.binding of Stx2B or IntC280 to the respective host receptors even in the presence of the two virulence factors together. Moreover,.the bispeci fi c neutralizer was relatively stable to harsh storage conditions and gastrointestinal pH extremes. Taking into account its.easy and economical production and superior pharmaceutical properties, we believe that a nanobody-based bispeci fi c neutralizer.would be more favorable and practical to be developed as a therapeutic to fi ght STEC in the developing world.