抗寨卡病毒包膜蛋白单克隆抗体的制备和鉴定

Preparation and characterization of monoclonal antibody against Zika virus envelope protein

目的制备抗寨卡病毒(zika virus,ZIKV)包膜蛋白(envelope protein,E蛋白)单克隆抗体,检测其免疫反应特异性。方法将ZIKV重组E蛋白免疫BALB/c小鼠,取小鼠脾细胞与骨髓瘤细胞用50%聚乙二醇(PEG)融合,经过含有次黄嘌呤、氨基喋呤和胸腺嘧啶的培养基筛选出阳性克隆,采取有限稀释法获得单克隆细胞株。利用筛选到的单抗作为一抗,行间接免疫荧光实验和免疫转印试验,鉴定单抗的反应特异性。结果筛选出1株针对寨卡病毒E蛋白的单克隆抗体(命名为13E7-E9),间接免疫荧光实验和免疫转印试验结果表明,该单抗可与寨卡病毒E蛋白结合,且与登革病毒无交叉。结论成功筛选1株抗寨卡病毒E蛋白的特异性单克隆抗体,可为研发寨卡病毒的免疫诊断试剂奠定基础。

Objective To screen and characterize monoclonal antibodies(mAbs)against the envelope protein of Zika virus(ZIKV E protein);to test its’s immune response specificity.Methods The SPF BALB/c mice were immunized with recombinant ZIKV E protein and the spleen cells were fused with myeloma cells using 50%polyethylene glycol.The positive cell clones were selected by medium containing hypoxanthine,aminopterin and thymine(HAT).The limited dilution method was used to obtain the single positive cell clone.The indirect immunofluorescence method(IFA)and western blot were established to characterize the mAb’s specificity.Results One hybridoma cell line called 13 E7-E9 with ZIKV E binding specificity was screened out,IFA showed the mAb specific interacted with ZIKV E protein with no cross-reactivity with dengue virus.Conclusion A specific monoclonal antibody against ZIKV E protein was successfully screened out,which can lay a foundation for the development of immunodiagnostic reagents for Zika virus.