番茄溃疡病菌单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibodies against the bacterial canker of tomato

作　　者：孔君[1,2] 刘箐 韩跃武[2] 刘金芝[3] 朱小清 席国雄 

机构地区：[1]甘肃出入境检验检疫局,甘肃兰州730020 [2]兰州大学基础医学院医学生物化学与分子生学研究所 [3]甘肃农业大学生命科学技术学院

出　　处：《植物检疫》2011年第4期42-46,共5页Plant Quarantine

基　　金：甘肃省科技厅中小企业创新基金计划(0802NCCA028);国家质检总局项目(2007IK259;2009IK276)

摘　　要：为制备并鉴定番茄溃疡病菌(Clavibacter michiganensis subsp.michiganensis,Cmm)的单克隆抗体(McAbs),用全菌皮下免疫BALB/C小鼠,采用B细胞杂交瘤技术,经免疫、融合、间接ELISA筛选和克隆等,获得稳定分泌抗体的阳性杂交瘤细胞株,得到了抗番茄溃疡病菌的单克隆抗体。经免疫后获得3株单抗分别为1A4、1C3和1B7,经亚类鉴定分别是IgM、IgG1、IgG1;纯化腹水间接ELISA效价分别为1:3.2×106、1:8.1×105、1:3.2×106;与其他同属不同亚种无交叉反应。结果表明:3株单克隆抗体均具有较高特异性和敏感性,可作为番茄溃疡病菌的检测抗体,其中,1A4的效果最好。番茄溃疡病菌单克隆抗体的获得为进一步研发番茄溃疡病检测试剂盒奠定了基础。To prepare and identify monoclonal antibodies（McAbs） agains Bacterial canker of tomato,Cmm,BALB/C mice were immunized with Cmm hypodermically and McAbs were gained by hybridoma cell lines secreting McAbs were obtained using routine hybridoma technology and ELISA screening.Three hybridoma cell lines specific to Cmm were obtained after screening with indirect ELISA and subcloning.They were designated as 1A4,1C3 and 1B7.Identification of the subclass indicated that the monoclonal antibody secreted by 1A4 cell line belonged to IgM,while those secreted by 1C3 and 1B7 belonged to IgG1.As demonst rated by indirect ELISA,the titers of the purified ascetic fluids of 1A4,1C3,1B7 were 1：3.2×106,1：8.1×105,1：3.2×106,respectively.The McAbs had no cross-reaction with some subspecies of bacteriums.It is evident that these monoclonal antibodies all show high degree of specificity and sensitivity,Which,1A4 was the best.The results show that the specific Cmm McAbs are developed and these McAbs may be useful in the development of detection assay.

关 键 词：单克隆抗体 番茄溃疡病菌 ELISA 

分 类 号：S436.412[农业科学—农业昆虫与害虫防治]