分析化学2012，Vol.40Issue(8)：1147-1152,6.DOI:10.3724/SP.J.1096.2012.20055
基于纳米金固定半抗原的电化学免疫传感器灵敏检测克伦特罗
Sensitive Electrochemical Immunosensor for Determination of Clenbuterol Using Gold Colloid-Mediated Hapten Immobilization
摘要
Abstract
An indirect competitive electrochemical immunosensor based on gold colloid-mediated hapten assembly was developed for the determination of clenbuterol (CLB), a β-agonist drug that has been proved to be very toxic to human beings. The sensing substrate was prepared using a gold electrode modified with a self-assembled monolayer of 1, 6-hexanedithiol that mediated the assembly of a gold colloid layer, which adsorbed clenbuterol and bovine serum albumin conjugates onto the electrode surface. After competition for the limited anti-CLB mouse monoclonal antibody between immobilized hapten and CLB analyte in sample solution, alkaline phosphatase (ALP)-labeled horse anti-mouse IgG antibody reacted selectively with the primary antibodies captured on the electrode surface. Electrochemical response was produced by the oxidation of enzymatic product of 1-naphthy1 phosphate. Under the optimal conditions, the ALP-mediated redox signals were inversely correlated to the concentration of the CLB analyte in the range of 0. 1μg/L to 1000μg/L. The regression equation was I(A) =8. 79 × 10-7 -2. 66×10-7 logC (μg/L) with a correlation coefficient of 0. 9960.The detection limit was estimated to be 20 pg/mL. Furthermore, the proposed electrochemical immunosensor was used to detect the content of clenbuterol in pork- tissues with an average relative standard of 7. 0%, and the recoveries were in the range from 89. 1% to 105. 6%. The statistical results for the same concentration level obtained by the proposed method and traditional indirect competitive ELISA were comparable, proving the feasibility of the electrochemical immunosensor for accurate determination of CLB in real samples.关键词
电化学免疫传感器/克伦特罗/纳米金组装/间接竞争免疫/1-萘酚磷酸酯Key words
Electrochemical immunosensor/ Clenbuterol/ Gold colloid assembly/ Indirect competitive assay/ 1-Naphthy1 phosphate引用本文复制引用
刘雪平,张一芳,吴会旺,沈国励,俞汝勤..基于纳米金固定半抗原的电化学免疫传感器灵敏检测克伦特罗[J].分析化学,2012,40(8):1147-1152,6.基金项目
本文系国家自然科学基金项目(Nos.21025521,21035001)和河南城建学院科学研究基金项目(No.2011JZD002)资助 （Nos.21025521,21035001）
