基于双层酶信号放大及纳米功能界面的微囊藻毒素电化学免疫传感器

引用本文: 尹艳艳, 侯丽, 张丽丽, 林学滨, 吴晓苹. 基于双层酶信号放大及纳米功能界面的微囊藻毒素电化学免疫传感器[J]. 分析化学, 2018, 46(4): 493-501. doi: 10.11895/j.issn.0253-3820.171055 shu

Citation: YIN Yan-Yan, HOU Li, ZHANG Li-Li, LIN Xue-Bin, WU Xiao-Ping. Electrochemical Immunosensor for Microcystin-LR Detection Based on Nano-composite Material Immobilization and Enzymatic Amplification[J]. Chinese Journal of Analytical Chemistry, 2018, 46(4): 493-501. doi: 10.11895/j.issn.0253-3820.171055 shu

基于双层酶信号放大及纳米功能界面的微囊藻毒素电化学免疫传感器

食品安全与生物分析教育部重点实验室(福州大学);福州大学化学学院, 福州 350108
基金项目:
本文系国家自然科学基金项目（No.21375019）和教育部创新团队（二期）项目（No.IRT15R11）资助

摘要: 设计了一种基于纳米复合膜双层酶信号放大的竞争型电化学免疫传感器，用于检测痕量微囊藻毒素-LR（Microcystin-（leucine-arginine），MCLR）。在多壁碳纳米管修饰玻碳电极上电沉积金纳米粒子形成纳米复合膜，作为MCLR抗体（anti-MCLR）和辣根过氧化物酶（HRP）的固定化载体；引入HRP作为封闭剂或者通过抗体捕获HRP标记的MCLR（HRP-MCLR），起封闭活性位点及协同催化的作用。利用MCLR与HRP-MCLR对纳米复合膜所固载抗体活性位点的竞争结合模式，采用微分脉冲伏安法（DPV）测定电极界面上HRP酶催化过氧化氢（H₂O₂）氧化对苯二酚（HQ）产生的还原电流，实现MCLR的定量检测。此传感器具有良好的特异性、稳定性与灵敏度，线性检测范围为0.1~100.0 μg/L，检出限为0.038 μg/L（S/N=3），对实际水样的加标回收率为72.9%~117.3%。

关键词:

English

Electrochemical Immunosensor for Microcystin-LR Detection Based on Nano-composite Material Immobilization and Enzymatic Amplification

Key Laboratory of Analytical Science of Food Safety and Biology(Ministry of Education & Fujian Province), College of Chemistry, Fuzhou University, Fuzhou 350108, China
Received Date: 01 July 2017
Revised Date: 25 December 2017
Fund Project:
This work was supported by the National Natural Science Foundation of China (No.21375019) and the Ministry of Education Innovation Team Development Plan (Phase Ⅱ) (No.IRT15R11).

Abstract: A competitive electrochemical immunosensor based on the nano-composite material immobilization and enzymatic amplification was designed for detection of microcystin-LR. Gold nanoparticles/carboxylated multi-walled carbon nanotubes (AuNPs/c-MWCNTs) composite film, which formed by electrodepositing of AuNPs on the C-MWCNTs modified glassy carbon electrode, was used for the immobilization of the antibody of microcystin-LR (anti-MCLR). Horseradish peroxidase (HRP) was introduced onto the nanocomposite interface by HRP blocked sensing interface and specific capture of antibody with target. It could be employed to catalyze the reduction of H₂O₂, and to block the possible remaining active sites as well. A competitive immunoreaction between antigen and MCLR-HRP was used for target analysis. In the presence of H₂O₂ and hydroquinone (HQ), MCLR could be indirectly detected with differential pulse voltammetry (DPV) method by determining the reduction current of HQ. Under the optimal conditions, the proposed immunosensor exhibited wide linear ranges in the concentration ranges of 0.1-100.0 μg/L, with a detection limit of 0.038 μg/L (S/N=3, n=8). The immunosensor showed good specificity, stability and sensitivity. It was used to determine MCLR in real water samples with the recoveries of 72.9%-117.3%.

Key words:

1. [1]
  Codd G A, Morrison L F, Metcalf J S. Toxicol. Appl. Pharmacol.,2005,203(3):264-272
2. [2]
  Lürling M, Faassen E J. Water Res.,2012,46(5):1447-1459
3. [3]
  Chen J, Xie P. Environ. Toxicol.,2005,20(6):572-584
4. [4]
  WHO. Guidelines for Drinking-Water Quality, 2rd ED, Geneva:World Health Organization,1997
5. [5]
  Shamsollahi H R, Alimohammadi M, Nabizadeh R, Nazmara S, Mahvi A. Glob. J. Health Sci.,2015,7(2):66-70
6. [6]
  Neffling M R, Spoof L, Meriluoto J. Anal. Chim. Acta,2009,653(2):234-241
7. [7]
  Liu L Q, Xing C G, Yan H J, Kuang H, Xu C L. Sensors,2014,14(8):14672-14685
8. [8]
  Garibo D, Flores C, Ceto X, Prieto-Simon B, Del Valle M, Caixach J,Diogène J, Campàs M. Environ. Sci. Pollut. Res.,2014,21(18):10652-10660
9. [9]
  Kimmel D W, Leblanc G, Meschievitz M E, Cliffel D E. Anal. Chem.,2012,84(2):685-707
10. [10]
  Sassolas A, Blum L J, Leca-Bouvier B D. Biotechnol. Adv.,2012,30(3):489-511
11. [11]
  Huo X H, Liu X Q, Liu J, Sukumaran P, Alwarappan S, Wong D K Y. Electroanalysis,2016,28(8):1730-1749
12. [12]
  Tang J, Tang D P. Microchim. Acta,2015,182(13):2077-2089
13. [13]
  Ansari S A, Husain Q. Biotechnol. Adv.,2012,30(3):512-523
14. [14]
  Nasir M, Nawaz M H, Latif U,Yaqub M, Hayat A, Rahim A. Microchim. Acta,2016:1-20
15. [15]
  Chen A C, Chatterjee S. Chem. Soc. Rev.,2013,42(12):5425-5438
16. [16]
  Wang J. Microchim. Acta,2012,177(3):245-270
17. [17]
  Saha K, Agasti S S, Kim C, Li X N, Rotello V M. Chem. Rev.,2012,112(5):2739-2779
18. [18]
  Shi X H, Gu W, Li B Y, Chen N N, Zhao K, Xian Y Z. Microchim. Acta,2014,181(1):1-22
19. [19]
  Tiwari J N, Vij V, Kemp K C, Kim K S. ACS Nano,2016,10(1):46-80
20. [20]
  Zhang J, Lei J P, Xu C L, Ding L, Ju H X. Anal. Chem.,2010,82(3):1117-1122
21. [21]
  Reverté L, Prieto-simón B, Campàs M. Anal. Chim. Acta,2016,908:8-21
22. [22]
  Zhao H M, Tian J P, Quan X. Colloids Surf. B,2013,103(1):38-44
23. [23]
  Ge S G, Liu W Y, Ge L, Yan M, Yan J X, Huang J D, Yu J H. Biosens. Bioelectron.,2013,49(35):111-117
24. [24]
  Hou L, Ding Y H, Zhang L L, Guo Y, Li M S, Chen Z, Wu X P. Sens. Actuators B,2016,233:63-70
25. [25]
  Li Y, Yuan R, Chai Y Q, Song Z J. Electrochim. Acta,2011,56(19):6715-6721
26. [26]
  Li W J, Yuan R, Chai Y Q, Chen S H. Biosens. Bioelectron.,2010,25(11):2548-2552
27. [27]
  Andrieux C P, Limoges B, Saveant J M, Yazidi D. Langmuir,2006,22(25):10807-10815
28. [28]
  Xu W J, Wu Y M, Yi H Y, Bai L J, Chai Y Q, Yuan R. Chem. Commun.,2014,50(12):1451-1453
29. [29]
  ZHANG Xin-Ai, SHEN Jian-Zhong, ZHANG Fan, MA Hai-Le, HAN En, DONG Xiao-Ya. Chinese J. Anal. Chem.,2014,42(9):1301-1306 张新爱, 申建忠, 张帆, 马海乐, 韩恩, 董晓娅.分析化学,2014,42(9):1301-1306
30. [30]
  Yu H W, Lee J, Kim S, Nguyen G, Kim I. Anal. Bioanal. Chem.,2009,394:2173-2181

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沈阳化工大学材料科学与工程学院沈阳 110142

基于双层酶信号放大及纳米功能界面的微囊藻毒素电化学免疫传感器

English

Electrochemical Immunosensor for Microcystin-LR Detection Based on Nano-composite Material Immobilization and Enzymatic Amplification

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于双层酶信号放大及纳米功能界面的微囊藻毒素电化学免疫传感器

English

Electrochemical Immunosensor for Microcystin-LR Detection Based on Nano-composite Material Immobilization and Enzymatic Amplification

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

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CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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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？

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