中国兽医科学2017，Vol.47Issue(7)：831-837,7.DOI:10.16656/j.issn.1673-4696.2017.07.005
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
摘要
Abstract
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/O myeloma cell.A hybridoma cell line(1C7) which could stably secrete monoclonal antibodies(McAbs) against the purified recombinant FMDV O serotype VP1 protein(O-rVP1) was screened and identified by hybridoma technique.The number of 1C7 chromosome was higher than that of any parental cells.The ascites titer was 1∶105,and the concentration was 4.7 mg/mL.The subtype of McAb 1C7 was identified as IgG2b.The relative affinity index of McAb 1C7 was 1.5 mol/L.The McAb 1C7 could specifically react with O serotypeFMDV,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 positire 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 on 1C7 McAb.关键词
O型口蹄疫病毒/VP1蛋白/单克隆抗体/制备/鉴定Key words
foot-and-mouth disease virus O serotype/VP1 protein/monoclonal antibody/preparation/identification分类
农业科技引用本文复制引用
李方方,高岭,闫若潜,吴志明,班付国,马震原,赵雪丽,李勤楠,张海迪,张旭..抗O型口蹄疫病毒VP1蛋白单克隆抗体的制备及鉴定[J].中国兽医科学,2017,47(7):831-837,7.基金项目
河南省科技创新人才计划项目(豫科人组[2016]4号) （豫科人组[2016]4号）
