一种基于宽谱特异性抗体的β2-激动剂多残留电化学免疫传感器的研制

食品科学 ›› 0, Vol. ›› Issue (): 0-0.

• 安全检测 • 下一篇

一种基于宽谱特异性抗体的β2-激动剂多残留电化学免疫传感器的研制

任鹏飞¹,邵科峰¹,张洁²,张红琳³,赵波¹

1. 南京师范大学
2. 南京师范大学化学与材料科学学院
3. 江苏南京晓庄学院

收稿日期:2015-10-12 修回日期:2016-02-25 出版日期:2016-04-25 发布日期:2016-04-13
通讯作者: 赵波 E-mail:zbchem@126.com
基金资助:
江苏省高校自然科学研究重大项目;江苏省科技支撑计划项目-社会发展;江苏省农业科技自主创新资金项目

Development of a multi-residue electrochemical immunoassay based on broad-specific antibody for determination of β2-agonists

Received:2015-10-12 Revised:2016-02-25 Online:2016-04-25 Published:2016-04-13
Contact: Bo ZHAO E-mail:zbchem@126.com

摘要/Abstract

摘要： 制备了沙丁胺醇-莱克多巴胺-牛血清蛋白偶联物（SAL-RAC-BSA）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（anti-SAL-RAC-BSA Abs），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（CV）和交流阻抗法（EIS）对电极的电化学性质进行了研究。采用间接竞争法进行了克伦特罗（CL）、沙丁胺醇（SAL）和特步他林（TB）三种β2-激动剂的同时检测，检出限依次为0.2、0.3和0.3ng/mL，线性范围依次为50~7000ng/mL、1~1500ng/mL和100~6000ng/mL。将传感器应用于猪肉、脂肪和猪肝三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β2-激动剂的高灵敏多残留免疫检测。

关键词: 宽谱特异性抗体, β2-激动剂, 多残留, 电化学免疫传感器

Abstract: Salbutamol-ractopamine-bovine serum albumin conjugate (SAL-RAC-BSA) was prepared and served as immunogen to obtain a broad-specific antibody (anti-SAL-RAC-BSA Abs) through rabbits immunization. Electrochemical immunosensor modified with reduced graphene oxide-chitosan composite (RGO-CTS) and SAL was constructed and the electrochemical behavior was studied by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Indirect competitive electrochemical immunoassay was carried out for the simultaneous determination of three β2-agonis, including clenbuterol (CL), salbutamol and terbutaline (TB). The proposed method indicated that the linear range was from 50 to 7000 ng/mL, 1 to 1500 ng/mL and 100 to 6000 ng/mL, respectively, while the detection limit was 0.2, 0.3 and 0.3 ng/mL, respectively. When the immunosensor was used for the detection of clenbuterol in spiked pork, fat and liver samples, the recovery rates range from 84.7% to 106%. In summary, a sensitive, rapid and simple electrochemical immunoassay was successfully developed for the determination of three β2-agonists.

Key words: broad-specific antibodies, β2-agonists, multi-residue, electrochemical immunosensor

中图分类号:

TS202.3

任鹏飞邵科峰张洁张红琳赵波. 一种基于宽谱特异性抗体的β2-激动剂多残留电化学免疫传感器的研制[J]. 食品科学, 0, (): 0-0.

参考文献

[1]张桂英.瘦肉精”的危害及监管探讨[J].中国动物检疫, 2011, 28(05):10-11 [2]庞苏纳, 于芳.“瘦肉精”及其危害简介[J].新疆畜牧业, 2009, (03):17-17 [3]Dominguez-Romero J C, Garcia-Reyes J F, Martinez-Romero R, et al.Dominguez-Romero J C, Garcia-Reyes J F, Martinez-Romero R, et al. Detection of main urinary metabolites of beta(2)-agonists clenbuterol, salbutamol and terbutaline by liquid chromatography high resolution mass spectrometry[J].Journal of Chromatography B, 2013, 923:128-135 [4]Blanca J, Munoz P, Morgado M, et al.Determination of clenbuterol,ractopamine and zilpaterol in liver and urine by liquid chromatography tandem mass spectrometry[J].Analytica Chimica Acta, 2005, 529(1-2):199-205 [5]Nielen M W F, Lasaroms J J P, Essers M L, et al.Multiresidue analysis of beta-agonists in bovine and porcine urine,feed and hair using liquid chromatography electrospray ionisation tandem mass spectrometry[J].Analytical And Bioanalytical Chemistry, 2008, 391(1):199-210 [6]Li Cun, Wu Yinliang, Yang Ting, et al.Simultaneous determination of clenbuterol,salbutamol and ractopamine in milk by reversed-phase liquid chromatography tandem mass spectrometry with isotope dilution[J].Journal Of Chromatography A, 2010, 1217(50):7873-7877 [7]Lin Xiaoyun, Ni Yongnian, Li Shuzhen, et al.A novel method for simultaneous analysis of three beta(2)-agonists in foods with the use of a gold-nanoparticle modified glassy carbon electrode and chemometrics[J].Analyst, 2012, 137(9):2086-2094 [8]冯亚娟, 牛司朋, 符雪文, 等.基于铂镍铁金属纳米花颗粒碳纳米片复合材料的克伦特罗电化学传感器的研制[J].分析化学, 2013, 41(03):427-431 [9]齐玉冰, 刘瑛, 宋启军.碳纳米管修饰电极分子印迹传感器快速测定沙丁胺醇[J].分析化学, 2011, 39(07):1053-1057 [10]袁利鹏, 杨金易, 徐振林, 等.荧光偏振免疫分析法检测沙丁胺醇[J].食品科学, 2013, 34(16):139-142 [11]陈小锋, 李瑞芳, 刘曙照.对克伦特罗具有高特异性的酶联免疫吸附分析法研究[J].分析化学, 2013, 41(6):940-943 [12]孙海新, 凌红丽, 张玉兰, 等.特布他林残留酶联免疫检测方法的建立[J].食品科学, 2010, 31(12):169-173 [13]Shen Wenjun, Zhuo Ying, Chai Yaqin, et al.Enzyme-Free Electrochemical Immunosensor Based on Host-Guest Nanonets Catalyzing Amplification for Procalcitonin Detection[J].Acs Applied Materials & Interfaces, 2015, 7(7):4127-4134 [14]Chen Xiaojun, Wang Yuanyuan, Zhou Jinjun, et al.Electrochemical impedance immunosensor based on three-dimensionally ordered macroporous gold film[J].Analytical Chemistry, 2008, 80(6):2133-2140 [15]Fang Yishan, Wang Haiying, Wang Lishi, et al.Electrochemical immunoassay for procalcitonin antigen detection based on signal amplification strategy of multiple nanocomposites[J].Biosensors & Bioelectronics, 2014, 51:310-316 [16]Feng Rui, Zhang Yong, Ma Hongmin, et al.Ultrasensitive non-enzymatic and non-mediator electrochemical biosensor using nitrogen-doped graphene sheets for signal amplification and nanoporous alloy as carrier[J].Electrochimica Acta, 2013, 97:105-111 [17]Bai Jing, Lai Yanjun, Jiang Dawei, et al.Ultrasensitive electrochemical immunoassay based on graphene oxide-Ag composites for rapid determination of clenbuterol[J].Analyst, 2012, 137(18):4349-4355 [18]Ambrosi A, Pumera M.Nanographite Impurities Dominate Electrochemistry of Carbon Nanotubes[J].Chemistry-A European Journal, 2010, 16(36):10946-10949 [19]Liu Yan, Liu Yang, Feng Hongbin, et al.Layer-by-layer assembly of chemical reduced graphene and carbon nanotubes for sensitive electrochemical immunoassay[J].Biosensors & Bioelectronics, 2012, 35(1):63-68 [20]Terrones M, Botello-Mendez A R, Campos-Delgado J, et al.Graphene and graphite nanoribbons: Morphology,properties,synthesis,defects and applications[J].Nano Today, 2010, 5(4):351-372 [21]Hummers W S, Offeman R E.Preparation of graphitic oxide[J].Journal of the American Chemical Society, 1958, 80(6):1339-1339 [22]邵科峰, 陈昌云, 颜妍, 等.基于石墨烯-壳聚糖修饰电极的免疫传感器检测三聚氰胺[J].食品科学, 2013, 34(16):221-225 [23]Meyer J C, Geim A K, Katsnelson M I, et al.The structure of suspended graphene sheets[J].Nature, 2007, 446(7131):60-63 [24]Lin Kuochiang, Hong Chuenpon, Chen Shenming.Simultaneous determination for toxic ractopamine and salbutamol in pork sample using hybrid carbon nanotubes[J].Sensors And Actuators B-Chemical, 2013, 177:428-436

一种基于宽谱特异性抗体的β2-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-specific antibody for determination of β2-agonists

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	刘凤银，苏珮韵，林浩标，刘汇，陈嘉仪，卢梓程，穆洪涛，李攻科. 联苯四氮唑沙坦类药物广谱性抗体的制备及其在降压类保健食品中多残留检测ic-ELISA法的应用[J]. 食品科学, 2023, 44(16): 331-339.
[2]	颜朦朦，李慧冬，张文君，陈子雷，郭长英，朱超，佘永新. 基于表面增强拉曼光谱标记技术的生物传感器用于农药残留检测的研究进展[J]. 食品科学, 2022, 43(17): 344-352.
[3]	杨倩雯，刘晓风，曾海娟，王金斌. 基于纳米材料的电化学免疫传感器及其在蛋白质检测中的研究进展[J]. 食品科学, 2021, 42(21): 347-354.
[4]	吴颖，范丛丛，苏晓娜，沈玉栋，谭庶，钟翠丽，许小炫，曾道平，杨金易. 基于智能手机同时检测4 种兽药残留的免疫芯片技术[J]. 食品科学, 2021, 42(2): 278-282.
[5]	刘凤银，李滢，王小鹏，吴晓丹，陈凤燕，梁岳，袁学文，穆洪涛. 酶抑制法快速检测牛奶中β-内酰胺酶抑制剂多残留[J]. 食品科学, 2021, 42(14): 248-254.
[6]	舒晓，褚能明，张雪梅，孟霞，李典晏，黄程兰，向嘉，杨俊英. 脂质去除分散固相萃取-气相色谱-串联质谱测定鸡蛋中62 种农药残留[J]. 食品科学, 2021, 42(14): 320-327.
[7]	刘希雅，田春妹，郑鹭飞，钟平胜，张馨方，任佳丽. 磁性三维还原石墨烯的制备及其在电化学免疫检测牛乳中结核杆菌H37Ra的应用[J]. 食品科学, 2020, 41(22): 308-314.
[8]	刘卫华，杨茜，刘敏轩，于文龙，王向红. 动物性食品中氟苯尼考检测的电化学免疫传感器构建[J]. 食品科学, 2020, 41(20): 307-313.
[9]	郝杰，姜洁，余建龙，路勇，毛婷，孙晓冬，杨丽梅. 固相萃取-超高效液相色谱-串联质谱法同时测定动物源性食品中多种兽药残留[J]. 食品科学, 2017, 38(12): 266-272.
[10]	任鹏飞，邵科峰，张洁，张红琳，赵波. 基于宽谱特异性抗体的β2-激动剂多残留电化学免疫传感器的研制[J]. 食品科学, 2016, 37(8): 236-241.
[11]	褚能明，孟霞，康月琼，唐偲雨，杨俊英. 分散固相萃取-气相色谱-串联质谱法测定茉莉花茶中86种农药残留[J]. 食品科学, 2016, 37(24): 239-246.
[12]	钟文英，王兴如，许丹科，李钟卉，李周敏. 可视化蛋白芯片法同时检测牛乳中残留的磺胺类和喹诺酮类药物[J]. 食品科学, 2016, 37(2): 193-197.
[13]	王锋，王宇，王弘，肖治理. 杂交-杂交瘤技术制备BsMAb的研究进展及其在食品安全检测中的应用[J]. 食品科学, 2016, 37(13): 251-256.
[14]	张毅，岳振峰，郭文，吴绍精，沈金灿，肖陈贵，林黎，华红慧，侯乐锡，易冰清. 动物源性食品中药物多残留分析的研究进展[J]. 食品科学, 2016, 37(1): 213-221.
[15]	樊晓博，谢兰心. 酶标抗原直接竞争ELISA检测食品中氟喹诺酮类药物多残留[J]. 食品科学, 2015, 36(24): 265-269.