抗微囊藻毒素单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibody against microcystin

作　　者：胡乐琴[1] 吴春燕[1] 张洋[1] 何培民[1] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306

出　　处：《生态科学》2014年第5期915-919,共5页Ecological Science

基　　金：上海市教育委员会科研创新项目(14YZ122)

摘　　要：构建了微囊藻毒素(Microcystin-LR,MC-LR)完全免疫原MCLR-KLH,免疫BALB/c小鼠,采用杂交瘤技术制备抗MCLR单克隆抗体,用间接竞争ELISA方法筛选抗体。经过3次克隆,获得2株可稳定分泌抗MCLR单克隆抗体的杂交瘤细胞株,腹水效价达1:8?104;IC50为0.81 ng?m L?1,最低检测限0.10 ng?m L?1,亲和常数3.8?108 L?mol?1,电泳结果显示抗体由1条重链和1条轻链组成,与MC-LR同系物MC-RR、MC-YR有较强的交叉反应,与MC-LW有弱交叉性反应,与河豚毒素无交叉反应。实验结果表明该抗体质量较好,将在微囊藻毒素的快速检测、产生机理等方面具有较大的科研和应用价值。A hybridoma cell line secreting monoclonal antibody against Microcystin-LR（MC-LR） was established after immunization of BALB/c mice with artificially synthesized MC-LR-KLH as antigen. The stimulated splenocytes were fused with SP2/0 myeloma cells to produce hybridomas. Two hybridoma cell lines were identified that secretes monoclonal antibody against Microcystin, after being subcloned for 3 cycles by indirect ELISA, which had an ascitic ELISA titre up to 640000. And the 50% inhibitory concentration of detection of was（0.81±0.05） μg·L-1 as measured by the competitive inhibition enzyme-linked immunosorbent assay（ci ELISA）. The Mc Ab will be quite useful for the determination of MC-LR residues.

关 键 词：微囊藻毒素 单克隆抗体 间接竞争ELISA(ciELISA) 

分 类 号：Q14[生物学—生态学]