免标记电化学发光免疫法传感器检测饲料中抗菌肽  

Label-free electrochemiluminescence immunosensor for determination of antibiotic peptide

作　　者：栾崇林[1] 蒋晓华[1] 代建国[1] 

机构地区：[1]深圳职业技术学院应用化学与生物技术学院,深圳518055

出　　处：《分析试验室》2016年第5期545-548,共4页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(21175037);广东省自然科学基金(S2011020005160);深圳市科技研发资金项目(JCYJ-20120617135709296)资助

摘　　要：构建了测定抗菌肽的免标记电化学发光免疫传感器。采用循环伏安法在金电极表面电聚合L-半胱氨酸（Cys）层,然后采用滴加的方式,通过Au-S共价和静电吸附作用再修饰纳米金颗粒于Cys层,进一步通过静电吸附作用固定抗菌肽抗体（例如天蚕素B抗体）,最后以牛血清白蛋白封闭非特异性吸附位点,制得目标传感器。将目标传感器在含有天蚕素B的溶液中于37℃温育1.5 h,由于抗原与目标传感器上的抗体会发生特异性的免疫反应,产生的免疫复合物会导致0.1 mol/L K2S2O8-0.1 mol/L PBS体系的ECL降低。在优化的条件下,天蚕素B在0.1-10 ng/m L范围内,ECL和天蚕素B质量浓度对数呈线性关系,线性方程为y=1352.0-520.54lgρ,r=0.996,检出限为30 pg/m L（S/N=3）。A label-free electrochemiluminescence（ ECL） immunosensor was prepared for the determination of antibiotic peptide. L-cysteine（ Cys） was electropolymerized on a bare gold electrode surface by cyclic voltammetry and then gold nanoparticles（ Au NPs） were modified on the layer of Cys by dropping through Au-S covalent bond and electrostatic adsorption. Subsequently,antibiotic peptide antibody（ e. g. cecropin B antibody）was immobilized on Au NPs through adsorption. Finally,bovine serum albumin（ BSA） was used to block the remaining sites on the surface to avoid non-specific adsorption. The immunosensor for antibiotic peptide assay was obtained. After incubating the prepared electrode in a solution containing cecropin B for 1. 5 h at 37 ℃,the ECL signal decreased in a solution containing 0. 1 mol / L K_2S_2O_8- 0. 1 mol / L PBS due to the formation of the complex of cecropin B antigen and cecropin B antibody through the specific immunoreaction. Under the optimized conditions,the ECL intensity of the immunosensor decreased linearly with the logarithmic concentration of cecropin B in the range of 0. 1 - 10 ng / m L with a limit of detection of 30 pg / m L（ S / N = 3）. The linear equation is y = 1352. 0- 520. 54 lgx,r = 0. 996. This immunosensor had been tested for its specificity,stability and reproducibility,all of which were well-performed. The developed immunosensor was used to determine cecropin B antibody in fish feed samples with satisfactory results.

关 键 词：免标记 电化学发光 免疫传感器 抗菌肽 天蚕素B 

分 类 号：O657.1[理学—分析化学]