盐酸克伦特罗单克隆抗体的制备及检测方法的初步建立

Preparation and characterization of monoclonal antibody against clenbuterol

选择一种新型载体阳离子化牛血清白蛋白(cBSA)与半抗原盐酸克伦特罗(CL)进行偶联,鉴定后作为免疫原免疫Balb/c小鼠,同时以卵清白蛋白(OVA)与CL偶联制备筛选抗原。应用杂交瘤细胞融合技术筛选后获得了1株稳定分泌抗CL单克隆抗体的杂交瘤细胞株,命名为2H5。抗体的亚类为IgM,腹水及细胞上清间接ELISA效价分别为1:20480和1:320,通过竞争阻断实验证明其具有良好的特异性及敏感性,将半抗原经酶标后应用竞争ELISA方法初步建立了CL的检测方法,检测限可达1ng/mL,为进一步制备CL残留检测试剂盒打下了基础。

6-week-old female BALB/c mice were immunized with a new carrier that synthesized CL-cBSA （cationised bovine serum albumin）. By using the hybridoma technique, A monoclonal antibody against clenbuterol（CL） was established. The antibody ,designed 2H5, of subclasses IgM, was isolated. The ELISA titers of ascite and cell supernatant were 1：20 480 and 1：320, respectively. It has good specificity and sensitivity by a competitive inhibition enzyme-linked immunosorbent assay（ciELISA）. The hapten was linked with enzyme and determination method was established primarily that based on competitive ELISA. The limit of this method was lng/ml. The study will be useful for studying the immunoassay in the determination of CL residues.