O型口蹄疫泛亚毒株单克隆抗体的制备及抗原表位差异分析  

The Preparation of Monoclonal Antibody against FMDV Type O/PanAsia and Differential Recognition of Antigenic Epitopes

作　　者：鲁会军[1] 金宁一[1] 韩松[1] 郑敏[1] 王凯[1] 贾雷立[1] 尹革芬[1] 金扩世[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2005年第5期494-496,共3页Chinese Journal of Veterinary Science

基　　金：科技部十五攻关项目(2004BA519A-40-2)

摘　　要：用纯化的O型口蹄疫泛亚毒株免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合,经3次克隆和间接ELISA筛选,获得了ⅢA11、ⅢC3和ⅢF10 3株稳定分泌单克隆抗体的杂交瘤细胞株.通过间接ELISA测定,单抗效价为:细胞培养上清1:160～1:640,腹水为1:5×104～1:4×105;经ELISA法测定,3株单克隆抗体均与泛亚株VP1蛋白反应,而不与A型口蹄疫病毒VP1蛋白反应;单抗的亚类鉴定结果表明,ⅢA11和ⅢF10分泌的抗体为IgG1亚类,ⅢC3分泌的抗体为IgG2b亚类.单克隆抗体抗原识别位点分析结果表明,ⅢA11与另外2种单克隆抗体的识别位点不同,而ⅢC3和ⅢF10的识别位点相近.The BALB/c mice were immuned with FMDV Type O/PanAsia. The myeloma cell line SP2/0 was fused with the spleen of immuned Balb/c, and by three limiting dilution, three specific monoclonal antibodies were screened by indirect ELISA. And three monoclonal antibody all could react with the FMDV. Among them, ⅢC3 belonged to IgG2b, Ⅲ All and Ⅲ F10 to IgG1. Their titers in cell culture supernatant were 1：320, 1：160 and 1：640respectively, and titersinascites were 1：2 × 10^5, 1：5 × 10^4 and 1：4 × 10^5 respectively. The three McAbs were specific for different kinds of antigenic binding epitopes, and they could specifically bind with VP1 protein of type O/PanAsia, but not with VP1 protein of type A.

关 键 词：口蹄疫 O型 泛亚毒株 单克隆抗体 识别位点 

分 类 号：S852.65[农业科学—基础兽医学]