抗沙丁胺醇单克隆抗体制备与鉴定

Preparation and Identification of Monoclonal Antibodies against Salbutamol

[目的]制备特异性抗沙丁胺醇单克隆抗体,为其免疫学检测方法的建立奠定基础。[方法]用SAL-BSA抗原免疫BALB/c小鼠;使用细胞融合技术建立抗SAL的单克隆抗体(SAL mAb)杂交瘤细胞株;体内诱生腹水法制备SAL mAb,并鉴定其免疫学特性。[结果]筛选出2D9和1C4两株杂交瘤细胞,其细胞培养上清效价达1∶104,腹水效价达1∶106;免疫球蛋白亚型鉴定为IgG1;抗体对SAL的半数抑制浓度(IC50)为1.14 ng/ml;与沙丁胺醇和盐酸克伦特罗的交叉反应率(CR)分别为100.00%和26.09%,与其他化合物无交叉反应。[结论]获得了高效价、敏感且异性的抗SAL抗体,为沙丁胺醇残留的免疫学检测方法的建立奠定了基础。

[Objective] To prepare specific monoclonal antibodies against salbutamol and thus to provide the basis for the immunological detection.[Method] BALB/c mice were immunized with SAL-BSA.Hybridoma lines（SAL mAb） secreting monoclonal antibody against SAL were established through cell fusion.Ascites of SAL mAb were prepared in vivo and the immunological characteristics were observed.[Result] Two hybridoma lines,2D9 and 1C4,were established,and the indirect ELISA titers of the cell supernatant and ascites were 1∶1×104 and 1∶1×106,respectively.Their isotype was IgG1.The median inhibitory concentration（IC50） of the SAL mAb against SAL was 1.14 ng/ml.It had cross-reactivity of 100.00% against salbutamol and 26.09% against clenobuterol hydrochloride,and no cross-reaction was found between the SAL mAb and other compounds.[Conclusion] The high-titer,sensitive and specific anti-SAL monoclonal antibodies have been produced,which makes it possible to develop immunoassay for detection of SAL residues.