猪流行性腹泻病毒N蛋白单克隆抗体的制备与鉴定

Preparation and Identification of Monoclonal Antibody against N Protein of Porcine Epidemic Diarrhea Virus

为制备猪流行性腹泻病毒(PEDV)单克隆抗体,采用生物信息学方法对CH/HNQX-3/14毒株N蛋白的抗原表位进行分析,将合成表位肽与载体偶联后免疫Balb/c小鼠,通过杂交瘤技术获得1株能稳定分泌抗PEDV单克隆抗体的杂交瘤细胞株(命名为9C4),其分泌抗体属于IgG2b亚类,轻链为κ链,腹水效价为1∶1.28×10^(4),能特异性识别PEDV。该单克隆抗体为研究PEDV N蛋白的功能和建立诊断方法提供了重要材料。

To prepare monoclonal antibody against PEDV,in this study,the antigenic epitopes of N protein from PEDV strainCH/HNQX-3/14 were analyzed using bioinformatics tools.Then the synthetic epitope peptides were coupled with keyhole limpethemocyanin(KLH)and used to immunize Balb/c mice.The monoclonal antibody(designated as mAb 9C4)against PEDV was obtained by hybridoma technique.The mAb belongs to IgG2b subclass and the light chain isκtype.The titer of ascites for 9C4 was 1∶1.28×10^(4) and could specifically recognize PEDV.This mAb would provide valuable tools for the study of PEDV N protein function and the establishment of PEDV diagnostic methods in the future.