Fu Lie, Jin Wei-Ping, Wang Wen-Hui, Wang Chen, Qian Sha-Sha, Wang Meng-Jun, Liu Rui-Lun, Li Song-Zhuang, Du Ya-Xin, Meng Sheng-Li, Guo Jing, Wang Ze-Jun, Chen Xiao-Qi, Shen Shuo
Wuhan Institute of Biological Products Co., Ltd.,, No.1 Huangjin Industrial Park Road, Jiangxia District, Wuhan, 430207, China.
Department of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China.
Virol J. 2024 Dec 30;21(1):338. doi: 10.1186/s12985-024-02596-4.
The hand, foot and mouth disease (HFMD) was caused by species of Enterovirus A and Enterovirus B in the Asian-Pacific region. Broad-spectrum monoclonal antibodies (mAb) that can bind multiple serotypes of enteroviruses have gradually become a research hotspot in the diagnosis, prevention and treatment of HFMD.
In this study, a mAb 1H4 was obtained using monoclonal antibody technology by immunizing purified virus particles of Coxsackievirus A5 (CV-A5). Examined by indirect immunofluorescence and Western blotting, 1H4 detected successfully all seven selected serotypes CV-A2, CV-A4, CV-A5, CV-A6, CV-A10, CV-A16 and EV-A71 of Enterovirus A and targeted structural protein VP1.
The mAb 1H4 showed no cross-reactivity to strains of Enterovirus B and Enterovirus C. A linear epitope WFYDGYPT was identified as the minimal binding region of 1H4 by indirect ELISAs with overlapped and truncated peptides of VP1. Alanine scanning test found that W, F, D, G, Y, P, and T were key residues in the epitope region. BLAST of the epitope in the NCBI genus Enterovirus protein database indicates that the epitope sequence is highly conserved among Enterovirus A species, but not among the other Enterovirus species.
The results suggest that the mAb 1H4 may be a useful tool for development with a cost-effective and accurate method for surveillance and early differentiation of serotypes from Enterovirus A species to other species.
在亚太地区，手足口病（HFMD）由肠道病毒A和肠道病毒B引起。能够结合多种肠道病毒血清型的广谱单克隆抗体（mAb）逐渐成为手足口病诊断、预防和治疗方面的研究热点。
在本研究中，通过用柯萨奇病毒A5（CV-A5）的纯化病毒颗粒免疫，利用单克隆抗体技术获得了单克隆抗体1H4。通过间接免疫荧光和蛋白质印迹法检测，1H4成功检测到了肠道病毒A的所有七种选定血清型CV-A2、CV-A4、CV-A5、CV-A6、CV-A10、CV-A16和EV-A71，并靶向结构蛋白VP1。
单克隆抗体1H4对肠道病毒B和肠道病毒C的菌株无交叉反应。通过用VP1的重叠肽和截短肽进行间接酶联免疫吸附测定（ELISA），确定线性表位WFYDGYPT为1H4的最小结合区域。丙氨酸扫描试验发现W、F、D、G、Y、P和T是表位区域的关键残基。在NCBI肠道病毒属蛋白质数据库中对该表位进行BLAST分析表明，该表位序列在肠道病毒A种之间高度保守，但在其他肠道病毒种之间不保守。
结果表明，单克隆抗体1H4可能是一种有用的工具，可用于开发一种经济高效且准确的方法，用于监测肠道病毒A种与其他种的血清型并进行早期区分。
