| 氟甲喹单克隆抗体制备、鉴定及间接竞争酶联免疫吸附分析法 |
| |
| 引用本文： | 王宇,沈玉栋,徐振林,雷红涛,王弘,孙远明.氟甲喹单克隆抗体制备、鉴定及间接竞争酶联免疫吸附分析法[J].分析化学,2010,38(3). |
| |
| 作者姓名： | 王宇 沈玉栋 徐振林 雷红涛 王弘 孙远明 |
| |
| 作者单位： | 广东省食品质量安全重点实验室,华南农业大学食品学院,广州,510642 |
| |
| 基金项目： | 国家自然科学基金(No.30901005);;国家863计划(No.2007AA10Z437);;广东省科技计划项目(No.2009A020101004);;广东省自然科学基金(No.06300421)项目资助 |
| |
| 摘 要： | 以氟甲喹(FLU)为原料,合成4个碳原子手臂的半抗原(FLUABA),采用活泼酯法与牛血清白蛋白(BSA)偶联制备免疫抗原,通过免疫Balb/c小鼠及细胞融合,获得1株稳定分泌抗氟甲喹单克隆抗体的杂交瘤细胞株DB6-E7,其抗体亚类为IgG1,亲和力常数(KA)为8.19×108L/mol。将氟甲喹、FLUABA及6个碳原子手臂的半抗原FLUACA分别与卵清白蛋白(OVA)偶联作为包被抗原,研究异源包被对间接竞争ELISA灵敏度的影响。结果表明,异源包被可显著提高ELISA方法的灵敏度。基于最佳异源包被(FLU-OVA)的酶联免疫吸附分析法的IC50为26.33μg/L,检出限为4μg/L,定量检测范围为8.0～114μg/L(IC20～IC80)。与喹诺酮类药物及结构类似物几乎不存在交叉反应,特异性高。此方法可满足畜禽产品中氟甲喹残留的快速筛查。
|
| 关 键 词： | 氟甲喹 单克隆抗体 异源包被 酶联免疫吸附分析法 |
Production and Identification of Monoclonal Antibody Against Flumequine and Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay |
| |
| WANG Yu,SHEN Yu-Dong,XU Zhen-Lin,LEI Hong-Tao,WANG Hong,SUN Yuan-Ming.Production and Identification of Monoclonal Antibody Against Flumequine and Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay[J].Chinese Journal of Analytical Chemistry,2010,38(3). |
| |
| Authors: | WANG Yu SHEN Yu-Dong XU Zhen-Lin LEI Hong-Tao WANG Hong SUN Yuan-Ming |
| |
| Institution: | Key Laboratory of Food Quality and Safety of Guangdong Province;College of Food Science;South China Agricultural University;Guangzhou 510642 |
| |
| Abstract: | The hapten of Flumequine( FLU) with four carbon atoms spacer arm( FLUABA) was synthesized and coupled to bovine serum albumin( BSA) as immunogen using activated ester method. Balb/c mice were immunized by the artificial immunogen and the splenocytes of immunized mice were fused with Sp2 /0 cells to obtain the monoclonal antibody( McAbs) . A hybridoma cell line( DB6-E7) secreting anti-flumequine McAbs was obtained by limited dilution method and screened by indirect enzyme-linked immunosorbent assay ( ELISA) ... |
| |
| Keywords: | Flumequine Monoclonal antibody Heterogenous antigen Enzyme-linked immunosorbent assay |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
