基于宽谱特异性抗体的β_2-激动剂多残留电化学免疫传感器的研制

Development of A Multi-Residue Electrochemical Immunoassay Based on Broad-Spectrum Specific Antibody for Determination of β_2-Agonists

制备了沙丁胺醇-莱克多巴胺-牛血清蛋白偶联物,并通过动物免疫获得了抗沙丁胺醇-莱克多巴胺-牛血清蛋白宽谱特异性抗体,同时制备了石墨烯-壳聚糖/沙丁胺醇修饰的电化学免疫传感器,并采用循环伏安法和交流阻抗法对电极的电化学性质进行研究。采用间接竞争法进行了克伦特罗、沙丁胺醇和特布他林3种β2-激动剂的同时检测,检出限依次为0.2、0.3 ng/m L和0.3 ng/m L,线性范围依次为50~7 000、1~1 500 ng/m L和100~6 000 ng/m L。将传感器应用于猪瘦肉、猪脂肪和猪肝3种实际样品中克伦特罗的加标检测,回收率为84.7%~106%。实现了β_2-激动剂的高灵敏多残留免疫检测。

Salbutamol-ractopamine-bovine serum albumin conjugate（SAL-RAC-BSA）was prepared and served as immunogen to obtain a broad-spectrum specific antibody（anti-SAL-RAC-BSA Abs）through immunization of rabbits.Electrochemical immunosensor modified with reduced graphene oxide-chitosan composite and SAL was constructed and its electrochemical behavior was studied by cyclic voltammetry and electrochemical impedance spectroscopy.Indirect competitive electrochemical immunoassay was carried out for the simultaneous determination of three β_2-agonis,including clenbuterol（CL）,salbutamol（SAL）and terbutaline（TB）.The proposed method indicated a linearity in the range from 50 to 7 000 ng/m L,1 to 1 500 ng/m L and 100 to 6 000 ng/m L for CL,SAL and TB,respectively,and the limits of detection were 0.2,0.3 and 0.3 ng/m L,respectively.The recovery rates of clenbuterol in spiked pork,fat and liver samples ranged from 84.7% to 106%.In summary,a sensitive,rapid and simple electrochemical immunoassay was successfully developed for the determination of three β_2-agonists.