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

引用本文: 贺锋, 王海洁, 李腾飞, 杜鹏飞, 王维婷, 王守经, 柳尧波, 马艳丽, 胡鹏, 谭天宇, 韩晴. 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[J]. 分析化学, 2023, 51(1): 102-111. doi: 10.19756/j.issn.0253-3820.221241 shu

Citation: 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. Magnetic Separation Immunosensor Based on Enzyme-linked Gold Nanocomposite Probe for Detection of Ractopamine[J]. Chinese Journal of Analytical Chemistry, 2023, 51(1): 102-111. doi: 10.19756/j.issn.0253-3820.221241 shu

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

贺锋 ^1,2, ,
王海洁 ^2,3, ,
李腾飞 ^{1,
,} ,
杜鹏飞 ^{2,
,} ,
王维婷 ^2, ,
王守经 ^2, ,
柳尧波 ^2, ,
马艳丽 ^2, ,
胡鹏 ^2, ,
谭天宇 ^1, ,
韩晴 ^1,2,

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

通讯作者: 李腾飞,E-mail:litengfei@hebeu.edu.cn; 杜鹏飞,E-mail:dupengfei2011@163.com

基金项目:
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)和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1.75pg/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。

关键词:

English

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

1.
School of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056038, China;
2.
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;
3.
College of Life Sciences, Yantai University, Yantai 264005, China

Corresponding author: LI Teng-Fei, ; DU Peng-Fei,

Received Date: 16 May 2022
Revised Date: 07 October 2022
Fund Project:
Supported by the Natural Science Foundation of Shandong Province, China (No.ZR2020QC250), the Agriculture Industry Technology System (No.CARS-38), the Modern Agricultural Technology Industry System of Shandong Province, China (No.SDAIT-10-10) and the Key Research and Development Projects of Hebei Province, China (No.22325501D).

Abstract: 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.

Key words:

1. [1]
  BRAMELD J M, PARR T. Proc. Nutr. Soc., 2016, 75(3):242-246.BRAMELD J M, PARR T. Proc. Nutr. Soc., 2016, 75(3):242-246.
2. [2]
  ZHANG W, WANG P L, SU X O. TrAC, Trends Anal. Chem., 2016, 85:1-16.ZHANG W, WANG P L, SU X O. TrAC, Trends Anal. Chem., 2016, 85:1-16.
3. [3]
  ZHAO Z, GU X, SU X O, LI J G, LI J, DONG Y C, YANG Y J, YAO T, QIN Y C. J. Anal. Toxicol., 2017, 41(1):60-64.ZHAO Z, GU X, SU X O, LI J G, LI J, DONG Y C, YANG Y J, YAO T, QIN Y C. J. Anal. Toxicol., 2017, 41(1):60-64.
4. [4]
  CHENG T Y D, SHELVER W L, HONG C C, MCCANN S E, DAVIS W, ZHANG Y L, AMBROSONE C B, SMITH D J. J. Agric. Food Chem., 2016, 64(40):7632-7639.CHENG T Y D, SHELVER W L, HONG C C, MCCANN S E, DAVIS W, ZHANG Y L, AMBROSONE C B, SMITH D J. J. Agric. Food Chem., 2016, 64(40):7632-7639.
5. [5]
  SLAVIN R E, YAEGER M J. Cardiovasc. Pathol., 2012, 21(4):334-338.SLAVIN R E, YAEGER M J. Cardiovasc. Pathol., 2012, 21(4):334-338.
6. [6]
  List of Drug Species Prohibited from Use in Feed and Animal Drinking Water (Ministry of Agriculture Announcement No. 176). 2019. 禁止在饲料和动物饮用水中使用的药物品种目录(农业部公告第176号). 2019.
7. [7]
  Ministry of Industry and Information Technology of the People's Republic of China. No. 41, The Ministry of Industry and Information Technology and Other 6 Departments Jointly Announced the Prohibition of the Production and Sale of Ractopamine. 2011. 中华人民共和国工业和信息化部.公告第41号, 工业和信息化部等6部门联合公告禁止生产和销售莱克多巴胺. 2011.
8. [8]
  PENG C Y, ZHANG S W, WU C L, FENG Y, ZHAO D, WANG X Q, BAI Z Y. Food Chem., 2021, 355:129662.PENG C Y, ZHANG S W, WU C L, FENG Y, ZHAO D, WANG X Q, BAI Z Y. Food Chem., 2021, 355:129662.
9. [9]
  XU J Q, XU S R, XIAO Y P, CHINGIN K, LU H Y, YAN R H, CHEN H W. Anal. Chem., 2017, 89(21):11252-11258.XU J Q, XU S R, XIAO Y P, CHINGIN K, LU H Y, YAN R H, CHEN H W. Anal. Chem., 2017, 89(21):11252-11258.
10. [10]
  VALESE A C, OLIVEIRA G A P, KLEEMANN C R, MOLOGNONI L, DAGUER H. J. Food Compos. Anal., 2016, 47:38-44.VALESE A C, OLIVEIRA G A P, KLEEMANN C R, MOLOGNONI L, DAGUER H. J. Food Compos. Anal., 2016, 47:38-44.
11. [11]
  HSIEH S Y, WANG C C, KOU H S, WU S M. J. Pharm. Biomed. Anal., 2017, 141:222-228.HSIEH S Y, WANG C C, KOU H S, WU S M. J. Pharm. Biomed. Anal., 2017, 141:222-228.
12. [12]
  HAN S J, ZHOU T J, YIN B J, HE P L. Microchim. Acta, 2018, 185(4):210.HAN S J, ZHOU T J, YIN B J, HE P L. Microchim. Acta, 2018, 185(4):210.
13. [13]
  DUAN N, GONG W H, WU S J, WANG Z P. Anal. Chim. Acta, 2017, 961:100-105.DUAN N, GONG W H, WU S J, WANG Z P. Anal. Chim. Acta, 2017, 961:100-105.
14. [14]
  XU M X, ZHAO W L, LIU J, HE T, HUANG J J, WANG J P. Anal. Lett., 2019, 52(11):1771-1787.XU M X, ZHAO W L, LIU J, HE T, HUANG J J, WANG J P. Anal. Lett., 2019, 52(11):1771-1787.
15. [15]
  HE L H, GUO C P, SONG Y P, ZHANG S, WANG M H, PENG D L, FANG S M, ZHANG Z H, LIU C S. Microchim. Acta, 2017, 184(8):2919-2924.HE L H, GUO C P, SONG Y P, ZHANG S, WANG M H, PENG D L, FANG S M, ZHANG Z H, LIU C S. Microchim. Acta, 2017, 184(8):2919-2924.
16. [16]
  SHELVER W L, SMITH D J. J. Immunoassay, 2000, 21(1):1-23.SHELVER W L, SMITH D J. J. Immunoassay, 2000, 21(1):1-23.
17. [17]
  SN/T 4818-2017. Determination of Ractopamine, Salbutamol, Clenbuterol Hydrochloride in Edible Animal for Imported and Export-Enzyme Linked Immunosorbent Assay Method. Industry Standard of Entry-exit Inspection and Quarantine of the People's Republic of China. 进出口食用动物中莱克多巴胺、沙丁胺醇、盐酸克伦特罗的测定酶联免疫吸附法.中华人民共和国出入境检验检疫行业标准. SN/T 4818-2017.
18. [18]
  LI G L, ZHANG X L, ZHENG F P, LIU J H, WU D. Food Chem., 2020, 332:127431.LI G L, ZHANG X L, ZHENG F P, LIU J H, WU D. Food Chem., 2020, 332:127431.
19. [19]
  DUAN N, LI C X, SONG M Q, WANG Z P, ZHU C Q, WU S J. Spectrochim. Acta, Part A, 2022, 265:120342.DUAN N, LI C X, SONG M Q, WANG Z P, ZHU C Q, WU S J. Spectrochim. Acta, Part A, 2022, 265:120342.
20. [20]
  YANG F F, XU L, DIAS A C P, ZHANG X Y. Food Chem., 2021, 350:129196.YANG F F, XU L, DIAS A C P, ZHANG X Y. Food Chem., 2021, 350:129196.
21. [21]
  GUO Q, HAN J J, SHAN S, LIU D F, WU S S, XIONG Y H, LAI W H. Biosens. Bioelectron., 2016, 86:990-995.GUO Q, HAN J J, SHAN S, LIU D F, WU S S, XIONG Y H, LAI W H. Biosens. Bioelectron., 2016, 86:990-995.
22. [22]
  CHEN S, ZHANG J B, GAN N, HU F T, LI T H, CAO Y T, PAN D D. Microchim. Acta, 2015, 182(3-4):815-822.CHEN S, ZHANG J B, GAN N, HU F T, LI T H, CAO Y T, PAN D D. Microchim. Acta, 2015, 182(3-4):815-822.
23. [23]
  WANG Y B, QI Q Q, ZHOU J R, LI H, FU L L. Food Control, 2020, 110:106989.WANG Y B, QI Q Q, ZHOU J R, LI H, FU L L. Food Control, 2020, 110:106989.
24. [24]
  YAN C, TENG J, LIU F Y, YAO B B, XU Z L, YAO L, CHEN W. Microchem. J., 2020, 159:105414.YAN C, TENG J, LIU F Y, YAO B B, XU Z L, YAO L, CHEN W. Microchem. J., 2020, 159:105414.
25. [25]
  DU P F, JIN M J, ZHANG C, CHEN G, CUI X Y, ZHANG Y D, ZHANG Y X, ZOU P, JIANG Z J, CAO X L, SHE Y X, JIN F, WANG J. Sens. Actuators, B, 2018, 256:457-464.DU P F, JIN M J, ZHANG C, CHEN G, CUI X Y, ZHANG Y D, ZHANG Y X, ZOU P, JIANG Z J, CAO X L, SHE Y X, JIN F, WANG J. Sens. Actuators, B, 2018, 256:457-464.
26. [26]
  DU P F, JIN M J, CHEN G, ZHANG C, CUI X Y, ZHANG Y D, ZHANG Y X, ZOU P, JIANG Z J, CAO X L, SHE Y X, JIN F, WANG J. Microchim. Acta, 2017, 184(10):3705-3712.DU P F, JIN M J, CHEN G, ZHANG C, CUI X Y, ZHANG Y D, ZHANG Y X, ZOU P, JIANG Z J, CAO X L, SHE Y X, JIN F, WANG J. Microchim. Acta, 2017, 184(10):3705-3712.
27. [27]
  GEORGANOPOULOU D G, CHANG L, NAM J M, THAXTON C S, MUFSON E J, KLEIN W L, MIRKIN C A. Proc. Natl. Acad. Sci. U. S. A., 2005, 102(7):2273-2276.GEORGANOPOULOU D G, CHANG L, NAM J M, THAXTON C S, MUFSON E J, KLEIN W L, MIRKIN C A. Proc. Natl. Acad. Sci. U. S. A., 2005, 102(7):2273-2276.
28. [28]
  ZHU J M, ZOU N L, ZHU D N, WANG J, JIN Q H, ZHAO J L, MAO H J. Clin. Chem., 2011, 57(12):1732-1738.ZHU J M, ZOU N L, ZHU D N, WANG J, JIN Q H, ZHAO J L, MAO H J. Clin. Chem., 2011, 57(12):1732-1738.

扫一扫看文章

计量

PDF下载量: 6
文章访问数: 318
HTML全文浏览量: 28

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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

通讯作者: 李腾飞,E-mail:litengfei@hebeu.edu.cn; 杜鹏飞,E-mail:dupengfei2011@163.com

English

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

Corresponding author: LI Teng-Fei, ; DU Peng-Fei,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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

通讯作者: 李腾飞,E-mail:litengfei@hebeu.edu.cn; 杜鹏飞,E-mail:dupengfei2011@163.com

English

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

Corresponding author: LI Teng-Fei, ; DU Peng-Fei,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs