沙丁胺醇单克隆抗体的制备及间接ELISA方法的建立  

Preparation of monoclonal antibody against salbutamol and establishment of indirect competitive ELISA

作　　者：彭述辉[1,2] 袁利鹏[2] 孙远明[2] 雷红涛[2] 王保玲[2] 徐振林[2] 庞杰[2] 

机构地区：[1]广州城市职业学院,广东广州510405 [2]广东省食品质量安全重点实验室/华南农业大学食品学院,广东广州510640

出　　处：《食品工业科技》2010年第7期171-175,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金(20877029;30700663);广东科技计划(Zgzhzd0808;2009B040500002;2009B080701068;2008B021400007);国家863计划(2007AA10Z437);国家"十一五"科技支撑计划(2006BAD27B02-05);福建省自然科学基金(2009J01061);福建省科技计划社会发展重点项目(2008Y0006)

摘　　要：制备抗沙丁胺醇单克隆抗体,鉴定其特异性并建立间接竞争ELISA检测方法。以合成的沙丁胺醇完全抗原免疫Balb/c小鼠,利用杂交瘤技术进行细胞融合,筛选克隆得到稳定分泌抗沙丁胺醇抗体的单克隆杂交瘤细胞,命名为4E4和3A2。经鉴定,两个抗体亚类都为IgG1,其腹水纯化后效价分别达到2.56×10^5和1.28×10^5。以效价高且特异性较好的4E4细胞株获得的抗体建立间接竞争ELISA方法,经条件优化后得到标准曲线,检测范围为4.76-84.99ng/mL,IC50为20.12ng/mL,检测限为3.25ng/mL,与克伦特罗交叉反应率为253.62%,与其它结构类似物没有明显交叉反应。本研究制备的单克隆抗体和建立的间接ELISA方法,可用于开发同时检测沙丁胺醇和克伦特罗的试剂盒。Monoclonal antibody against Salbutamol（SAL）was prepared and the specificity of the antibody was identified to develop an indirect competitive enzyme-linked immunosobent assay（ELISA）for SAL. Splenocytes from mice immunized with SAL -BSA were fused with SP2/0 myeloma cells,and hybridomas secreting antibodies against SAL were selected and cloned. Two stable monoclonal antibodies 4E4 and 3A2 of subclasses IgG1 were isolated and the titers of corresponding ascites were 2.56×10^5 and 1.28×10^5,respectively. Based on the antibody produced by cell 4E4,an indirect competitive ELISA（icELISA）was developed for the quantitative detection of SAL. The IC50 of standard curve was 20.12ng/mL and the limit of detection for SAL was 3.25ng/mL,with a linear ranged from 4.76 to 84.99ng/mL. The obtained SAL monoclonal antibody had 253.62% cross-reactivity（CR%）to Clenbuterol（CBL）and showed scarcely cross-reactivity with other structural analogs.The monoclonal antibody and the method was suitable for developing a commercial immunoassay kit for detecting the residue of SAL and CBL simultaneously.

关 键 词：沙丁胺醇 单克隆抗体 间接ELISA 

分 类 号：Q785[生物学—分子生物学]