本试验采用苦马豆素(swainsonine,SW)人工抗原SW-OVA免疫接种BALB/c小鼠,通过杂交瘤技术建立了1株能稳定分泌SW单克隆抗体的杂交瘤细胞1F10,杂交瘤细胞染色体数目为45~50对。经间接ELISA检测,该株细胞培养上清中抗体效价为1∶25 600,诱生腹水效价为1∶80 000。单克隆抗体的亚型为IgG1,亲和力解离常数为1.14×1010,腹水抗体纯化后纯度可达98%,回收率80%,SDS-PAGE凝胶电泳可见单克隆抗体的轻链、重链分子质量分别为25和50ku,Western blotting检测抗体能特异性的识别SW。其线性检测范围为4~128μg/mL(R2=0.9969),与BSA、明胶、多聚赖氨酸、α-甘露糖苷等无交叉反应。本试验制备的SW单克隆抗体为免疫学检测SW和免疫学防制家畜疯草中毒奠定基础。
In this test,BALB/c mice were immunized by SW-OVA for the preparation of monoclonal antibody against swainsonine(SW).Hybridoma cells that could secrete specific antibody against SW were prepared by hybridoma technique,and then the strain that secreted monoclonal antibody against SW designated 1F10 was prepared,and the chromosome average number of 1F10 cell was 45 to 50couples.The ELISA titers of cell supernatant were 1∶25 600,and that of ascites were 1∶80 000.The subclasses of monoclonal antibody was IgG1,and the affinity constant was 1.14×1010,the purity of ascites antibodies was up to 98%,and the recovery rate was 80%.The result of sodium dodecyl sulfate polyacrylamide gel electropheresis proved that purified antibody had been obtained that the molecular weight of H-chain and L-chain of antibody was about50 and 25ku.The monoclonal antibody against SW specifically bound to SW determined by Western blotting.The linear range was 4to 128μg/mL(R2=0.9969)and no cross-reactivity was detected with BSA,gelatin,polylysine,me-Gal,etc.The result laid the foundation for immunodetection on SW and immunological prevention of animal toxic disease.