基于酶联金纳米复合探针的磁分离免疫传感器检测莱克多巴胺  

Magnetic Separation Immunosensor Based on Enzyme-linked Gold Nanocomposite Probe for Detection of Ractopamine

作　　者：贺锋 王海洁 李腾飞 杜鹏飞 王维婷[2] 王守经[2] 柳尧波[2] 马艳丽 胡鹏[2] 谭天宇 韩晴 HE Feng;WANG Hai-Jie;LI Teng-Fei;DU Peng-Fei;WANG Wei-Ting;WANG Shou-Jing;LIU Yao-Bo;MA Yan-Li;HU Peng;TAN Tian-Yu;HAN Qing(School of Life Sciences and Food Engineering,Hebei University of Engineering,Handan 056038,China;Key Laboratory of Novel Food Resources Processing,Ministry of Agriculture,Shandong Provincial Key Laboratory of Agro-Products Processing Technology,Institute of Agro-Food Sciences and Technology,Shandong Academy of Agricultural Sciences,Jinan 250100,China;College of Life Sciences,Yantai University,Yantai 264005,China)

机构地区：[1]河北工程大学生命科学与食品工程学院,邯郸056038 [2]山东省农业科学院农产品加工与营养研究所,山东省农产品精深加工技术重点实验室,农业农村部新食品资源加工重点实验室,济南250100 [3]烟台大学生命科学学院,烟台264005

出　　处：《分析化学》2023年第1期102-111,共10页Chinese Journal of Analytical Chemistry

基　　金：山东省自然科学青年基金项目(No.ZR2020QC250);国家现代农业(肉羊)产业技术体系建设专项(No.CARS-38);山东省现代农业产业技术体系-羊创新团队建设项目(No.SDAIT-10-10);河北省重点研发计划项目(No.22325501D)资助。

摘　　要：构建了一种基于新型酶联金纳米复合探针(E-GNPs)的磁分离竞争免疫传感器,用于检测莱克多巴胺(Ractopamine,RAC)。通过链霉亲和素与生物素的特异性相互作用,在抗体上标记辣根过氧化物酶(Horseradish peroxidase,HRP),采用静电组装法将抗体修饰于金纳米颗粒表面得到E-GNPs,引入卵清蛋白(Ovalbumin,OVA)-RAC半抗原(Hapten)复合物包被的磁珠进行竞争反应,通过显色反应实现目标物的定量分析。当目标分子RAC浓度变化时,酶催化底物显色随之改变,并且RAC的浓度与显色信号强度呈线性关系。紫外-可见吸收光谱表征结果表明,一个E-GNPs可携带11个HRP分子,使得该免疫传感器具有较高的灵敏度,检出限低至1.75 pg/mL,较传统酶联免疫方法灵敏度提高了10倍。交叉反应实验结果表明,此传感器对RAC的选择性良好,可应用于猪肉、牛肉和羊肉等实际样品中RAC的检测,加标回收率为88.3%~103.4%。本方法为动物源食品中RAC的快速筛查提供了一种新思路。A magnetic separation competitive immunosensor based on novel enzyme-linked gold nanocomposite probes(E-GNPs)was constructed for detection of ractopamine(RAC).Through the specific interaction between streptavidin and biotin,horseradish peroxidase(HRP)was labeled on the antibody that was modified on the surface of gold nanoparticles(AuNPs)by electrostatic assembly method to obtain E-GNPs.Ovalbumin(OVA)-RAC hapten coated magnetic beads(MBs)was introduced for a competitive reaction,and quantitative analysis of the target was achieved by a color reaction.With the change of the concentration of target molecule RAC,the color of the enzyme-catalyzed substrate changed,and the concentration of RAC had a linear relationship with the color signal intensity.The UV-vis absorption spectroscopy characterization results showed that one E-GNP could carry 11 HRP molecules,which made the immunosensor highly sensitive.The prepared sensor was successfully used for detection of RAC with a detection limit of 1.75 pg/mL,showing 10 times higher sensitivity than that of traditional ELISA method.Cross-reaction experiments showed that the sensor had good selectivity for RAC,and could be applied to real samples such as pork,beef and mutton,with spiked recoveries ranging from 88.3%to 103.4%.This analytical method provided a new idea for rapid screening of RAC in animal-derived foods.

关 键 词：莱克多巴胺 金纳米颗粒 磁性微珠 辣根过氧化物酶 

分 类 号：TS207.3[轻工技术与工程—食品科学]