抗大田软海绵酸单克隆抗体的制备及免疫学特性分析

Preparation of monoclonal antibody against okadaic acid and analysis for its immunological traits

目的制备了赤潮毒素大田软海绵酸(okadaic acid,OA)单克隆抗体,并分析其免疫学特性。方法用人工抗原大田软海绵酸-小牛血清蛋白偶联物(OA-BSA)免疫Balb/c小鼠,采用杂交瘤技术经初筛、复筛和亚克隆,筛选到1株可稳定分泌抗OA单克隆抗体的杂交瘤细胞株3H4。对抗体效价、亚型、特异性、敏感性进行免疫学鉴定与分析。结果抗体亚型为IgG2b;腹水效价为1∶1.28×106;50%抑制质量浓度为2.852 ng/ml;最低检测限为0.45 ng/ml;电泳结果显示抗体,由1条重链和1条轻链组成;与OA同系物PTX1、GYM、SPT1、YTX交叉反应率为0。结论制备的OA单抗效价和特异性均较好,可用于OA的免疫学检测。

We aim to prepare monoclonal antibodies against okadaic acid（OA） and analyze its immunological traits.With synthesized OA-bovine serum albumin（OA-BSA） as antigen,Balb/c mice were immunized.Then by hybridoma technique,a hybridoma cell line 3H4 was established,which could stably secret monoclonal antibody against OA.The secreted antibody was analyzed for titer,haplotype,specificity and sensitiveness.The haplotype of the monoclonal antibody（McAb） was IgG2b;the indirect ELISA showed the titer of ascites was 1∶1.28×106,and the 50% inhibitory concentration was 2.852 μg/L measured by the competitive inhibition enzyme-linked immunosorbent assay（ciELISA）.The lowest detectable limit of the McAb was 0.45 ng/ml.SDS-PAGE indicated that the McAb protein was made up of a weight chain and a light chain.Furthermore,the McAb was quite specific as no cross-reactivity with PTX1,GYM,SPT1,YTX.From all the result,we presume that the McAb prepared in this study would be quite useful for the determination of OA.