抗沙丁胺醇单克隆抗体制备与鉴定 |
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引用本文: | 李春生,吴萌,李君华,齐颖颖,李玉静,闫静辉.抗沙丁胺醇单克隆抗体制备与鉴定[J].安徽农业科学,2011(10):5931-5932. |
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作者姓名: | 李春生 吴萌 李君华 齐颖颖 李玉静 闫静辉 |
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作者单位: | 河北省科学院生物研究所;河北交通职业技术学院 |
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基金项目: | 河北省科技支撑计划项目(09227134D);石家庄市科技支撑计划项目(10150142A) |
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摘 要: | 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。
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关 键 词: | 沙丁胺醇 杂交瘤 单克隆抗体 |
Preparation and Identification of Monoclonal Antibodies against Salbutamol |
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Institution: | LI Chun-sheng et al(Biology Institute,Hebei Academy of Science,Shijiazhuang,Hebei 050081) |
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Abstract: | 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. |
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Keywords: | Salbutamo Hybridomas Monoclonal antibody |
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