分泌抗芜菁花叶病毒株系特异性单克隆抗体杂交瘤细胞株的建立及抗体理化性质测定  

The Establishment of Hybridoma Cell Line Secreting Specific Monoclonal Antibodies Against Turnip Mosaic Virus and Analysis of Properties of the McAbs

作　　者：杨峰[1] 蔡少华[1] 徐玲[2] 冯兰香[2] 

机构地区：[1]中国农业科学院生物技术研究中心 [2]中国农业科学院蔬菜花卉研究所

出　　处：《生物工程学报》1992年第4期342-347,共6页Chinese Journal of Biotechnology

摘　　要：用经芜菁花叶病毒(TuMV)免疫的BALB/c小鼠脾细胞与骨髓瘤细胞(Sp2/0-Ag14)融合,经3次克隆化培养和ELISA筛选,建立了5类分泌抗TuMV的单克隆抗体(McAb)的杂交瘤细胞株。其中4类分别对TuMV-C1株系、TuMV-C3株系、TuMV-C4株系和TuMV-C5株系具有特异性反应,第5类对TuMV 5个株系均有反应。以双抗体夹心ELISA和间接ELISA检测,上述McAb同CaMV、CMV、TMV,PVX和PVY等均不产生交叉反应,小鼠腹水McAb的滴度在1:256000—2048000之间,多为1:1024000,比常规多抗血清高400倍左右。对上述杂交瘤细胞系和McAb的生物学特性及理化性质进行了鉴定。用SDS-PAGE、Westernblotting对TuMV外壳蛋白亚基、McAb识别位点进行了分析,并就McAb区分TuMV株系进行了讨论。Monoclonal antibodies of hybridomas derived from fusing the mouse spleencytes immunized by TuMV with BALB/c mouse myeloma cells (Sp2/0-Ag14). Five kinds of hybridoma cell line were produced by indirectELISA screening and cloning three times with Limiting dilution. Four kinds of hybridoma respectively produced antibodies reactive to TuMV C1,C3,C4,C5. One kind reactive to all five strains of TuMV. In indirect-ELISA and sandwich-ELISA tests, TuMV specific monoclonal antibodies didn't react with CaMV, CMV, TMV, PVX and PVY. Antibody titers of ascitic fluids were about 11256000 to 2048000 in indirect-ELISA. Biological property and physical and chemical property of the hybridoma cell lines and McAbs was identified. The identification of TuMV strains, the specificity and stability of McAbs, the coat proteins and antigenic site of TuMV were discussed in this paper witb SDS-PAGE and western-blotting.

关 键 词：芜菁花叶病毒 杂交瘤 单克隆抗体 

分 类 号：S188[农业科学—农业基础科学]