肠道病毒71型和柯萨奇病毒A16鼠源性广谱中和活性单克隆抗体的制备

Preparation of pan-neutralizing mouse monoclonal antibodies against enterovirus 71 and coxsackievirus A16

目的制备可以同时阻断肠道病毒71型(EV71)及柯萨奇病毒A组16型(CV-A16)感染的中和活性单克隆抗体(mAb)。方法采用EV71病毒体蛋白1(VP1)羧基端的163~177位氨基酸(SP55)免疫BALB/c小鼠,通过杂交瘤技术制备mAb,用EV71中和抗原表位SP55以及高度同源的CV-A16 VP1羧基端的第163~177位氨基酸(PEP55)同时进行检测,筛选同时与EV71和CV-A16发生交叉反应的mAb,并进行体外中和试验检测对EV71和CV-A16的中和作用,并分析mAb的生物学特性。结果筛选获得1株可以同时中和EV71及CV-A16的mAb 6E5,其重链为IgG1亚类,轻链为Kappa链;细胞微量中和实验表明其抗EV71感染的中和效价为1∶128,抗CV-A16感染的中和效价为1∶32。结论成功获得1株能同时中和EV71及CV-A16的广谱中和活性mAb。

Objective To prepare a neutralizing monoclonal antibody(mAb) that can simultaneously block enterovirus 71(EV71) and coxsackievirus A16(CV-A16) infections. Methods BALB/c mice were immunized with 163-177 amino acids(SP55) of the C-terminal of EV71 virion particle 1(VP1) protein, and the mAbs were prepared by hybridoma technology. Neutralization antigenic epitope SP55 of EV71 and the highly homologous CV-A16 VP1 protein C-terminal 163-177 amino acids(PEP55) were applied to detect the mAbs that cross-reacted with EV71 and CV-A16 at the same time, and an in vitro neutralization test was conducted to detect the neutralization effect of EV71 and CV-A16, and to analyze the biological characteristics of the mAb. Results A mAb 6E5 with IgG1 subclass heavy chain and Kappa light chain was prepared, 6E5 mAb can cross-neutralize both EV71 and CV-A16. The mAb 6E5 could neutralize EV71 with a titer of 1∶128, and CV-A16 with a titer of 1∶32. Conclusion We have prepared a mAb 6E5 with a pan-spectrum neutralizing activity that can neutralize EV71 and CV-A16 at the same time.