抗O型口蹄疫病毒VP1蛋白单克隆抗体的制备及鉴定  

Preparation and identification of monoclonal antibodies against VP1 protein of foot-and-mouth disease virus O serotype

作　　者：李方方[1] 闫若潜[2] 吴志明[3] 班付国[2] 马震原[2] 赵雪丽[2] 李勤楠 张海迪[1] 张旭[2] 高岭[2] LI Fang-fang YAN Ruo-qian WU Zhi-ming BAN Fu-guo MA Zhen-yuan ZHAO Xue-li LI Qin-nan ZHANG Hai-di ZHANG Xu GAO Ling(College ofA nimal Science and Technology,Henan University of Science and Technology,Luoyang 471003, China Henan Centre for A nimal Disease Control ＆ Prevention, Zhengzhou 450008, China He nan Centre for A nimal Products Quality Inspection, Zhengzhou 450008, China)

机构地区：[1]河南科技大学动物科技学院,河南洛阳471003 [2]河南省动物疫病预防控制中心,河南郑州450008 [3]河南省畜产品质量监测检验中心,河南郑州450008

出　　处：《中国兽医科学》2017年第7期831-837,共7页Chinese Veterinary Science

基　　金：河南省科技创新人才计划项目(豫科人组[2016]4号)

摘　　要：以高浓缩的O型口蹄疫病毒(FMDV)灭活疫苗为抗原免疫BALB/c小鼠,取其脾细胞与小鼠SP2/0骨髓瘤细胞进行融合,以重组表达的O型重组VP1蛋白(O-r VP1)为筛选抗原,利用杂交瘤技术获得1株可稳定分泌抗O型FMDV VP1蛋白抗体的杂交瘤细胞株1C7,染色体数高于任一亲本细胞的染色体数目;腹水的抗体效价为1︰105,质量浓度为4.7 mg/m L。1C7单克隆抗体为IgG2b亚型,相对亲和力常数为1.5 mol/L,能够与O型FMDV特异性地结合,且仅与结构蛋白VP1反应。稳定性鉴定结果表明1C7株能稳定分泌抗体。阻断ELISA试验表明1C7单克隆抗体能够被O型FMDV免疫阳性血清特异性地阻断。本研究为进一步建立O型FMDV抗体的阻断ELISA及胶体金免疫层析快速检测方法奠定了基础。Female BALB/c mice were immunized with high concentration of foot-and-mouth disease virus（FMDV） O serotype inactivated vaccine,and the spleen cells of immunized mice were fused with SP2/0myeloma cell.A hybridoma cell line（1C7） which could stably secrete monoclonal antibodies（McAbs） against the purified recombinant FMDV O serotype VP1 protein（O-r VP1） was screened and identified by hybridoma technique.The number of 1C7 chromosome was higher than that of any parental cells.The ascites titer was1∶10^5,and the concentration was 4.7 mg/m L.The subtype of McAb 1C7 was identified as IgG2 b.The relative affinity index of McAb 1C7 was 1.5 mol/L.The McAb 1C7 could specifically react with O serotype FMDV,but not cross reacted to A serotype,Asia 1 serotype FMDV as well as swine vesicular disease virus.It could also react with VP1 protein but not cross reacted to VP2,VP3 and VP4 protein. The 1C7 was passaged to 30 generations,revived after frozen to 12 months in liquid nitrogen,and always secreted antibody stably.The blocking ELISA showed that McAb 1C7 could be specifically blocked by FMDV O serotype immune posi-tive serum.This study laid the foundation for detection of pathogens and antibodies of FMDV O serotype using the blocking ELISA and colloidal gold rapid detection of immune chromatography method based on1C7 McAb.

关 键 词：O型口蹄疫病毒 VP1蛋白 单克隆抗体 制备 鉴定 

分 类 号：S852.659.6[农业科学—基础兽医学]