一次性丝网印刷碳电极上辣根过氧化物酶的灵敏电化学检测
Sensitive electrochemical detection of horseradish peroxidase at disposable screen-printed carbon electrode.
作者信息
Lee Ai-Cheng, Liu Guodong, Heng Chew-Kiat, Tan Swee-Ngin, Lim Tit-Meng, Lin Yuehe
机构信息
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543.
出版信息
Electroanalysis. 2008 Aug 5;20(18):2040. doi: 10.1002/elan.200804287.
Abstract
A rapid, simple and sensitive electrochemical assay of horseradish peroxidase (HRP) performed on disposable screen-printed carbon electrode was developed. HRP activities were monitored by square-wave voltammetric (SWV) measuring the electroactive enzymatic product in the presence of o-aminophenol and hydrogen peroxide substrate solution. SWV analysis demonstrated a greater sensitivity and shorter analysis time than the widely used amperometric and differential-pulsed voltammetric methods. The voltammetric characteristics of substrate and enzymatic product as well as the parameters of SWV analysis were optimized. Under optimized conditions, a linear response for HRP from 0.003 - 0.1 U/mL and a detection limit of 0.002 U/mL (1.25×10(-15) mol in 25 μL) were obtained with a good precision (RSD = 8%; n = 6). This rapid and sensitive HRP assay with microliters-assay volume could be readily integrated to portable devices and point-of-care (POC) diagnosis applications.
摘要
开发了一种在一次性丝网印刷碳电极上对辣根过氧化物酶（HRP）进行快速、简单且灵敏的电化学检测方法。通过方波伏安法（SWV）在邻氨基酚和过氧化氢底物溶液存在的情况下测量电活性酶产物来监测HRP活性。与广泛使用的安培法和差分脉冲伏安法相比，SWV分析显示出更高的灵敏度和更短的分析时间。对底物和酶产物的伏安特性以及SWV分析参数进行了优化。在优化条件下，获得了HRP在0.003 - 0.1 U/mL范围内的线性响应以及0.002 U/mL（25 μL中为1.25×10(-15) mol）的检测限，精密度良好（RSD = 8%；n = 6）。这种具有微升检测体积的快速灵敏的HRP检测方法可以很容易地集成到便携式设备和即时检测（POC）诊断应用中。
相似文献
1
Sensitive electrochemical detection of horseradish peroxidase at disposable screen-printed carbon electrode.一次性丝网印刷碳电极上辣根过氧化物酶的灵敏电化学检测
Electroanalysis. 2008 Aug 5;20(18):2040. doi: 10.1002/elan.200804287.
2
Design and development of a highly stable hydrogen peroxide biosensor on screen printed carbon electrode based on horseradish peroxidase bound with gold nanoparticles in the matrix of chitosan.基于壳聚糖基质中与金纳米粒子结合的辣根过氧化物酶，在丝网印刷碳电极上设计和开发一种高度稳定的过氧化氢生物传感器。
Biosens Bioelectron. 2007 Apr 15;22(9-10):2071-8. doi: 10.1016/j.bios.2006.09.011. Epub 2006 Oct 13.
3
Novel and highly stable strategy for the development of microfluidic enzymatic assays based on the immobilization of horseradish peroxidase (HRP) into cotton threads.基于辣根过氧化物酶（HRP）固定到棉线中开发微流控酶分析的新颖且高度稳定的策略。
Talanta. 2023 Jan 15;252:123889. doi: 10.1016/j.talanta.2022.123889. Epub 2022 Aug 27.
4
Electrochemical detection of leukemia oncogenes using enzyme-loaded carbon nanotube labels.
Analyst. 2014 Sep 7;139(17):4223-30. doi: 10.1039/c3an01156a.
5
A amperometric biosensor for hydrogen peroxide by adsorption of horseradish peroxidase onto single-walled carbon nanotubes.基于辣根过氧化物酶在单壁碳纳米管上的吸附作用构建安培型过氧化氢生物传感器。
Colloids Surf B Biointerfaces. 2012 Feb 1;90:62-7. doi: 10.1016/j.colsurfb.2011.09.045. Epub 2011 Oct 4.
6
Evaluation of seven cosubstrates in the quantification of horseradish peroxidase enzyme by square wave voltammetry.采用方波伏安法评价七种共底物对辣根过氧化物酶的定量分析。
Talanta. 2012 Jan 15;88:468-76. doi: 10.1016/j.talanta.2011.11.016. Epub 2011 Nov 9.
7
Dual amplification strategy of highly sensitive thrombin amperometric aptasensor based on chitosan-Au nanocomposites.基于壳聚糖-金纳米复合材料的高灵敏度凝血酶安培适体传感器的双重放大策略。
Analyst. 2012 Aug 7;137(15):3488-95. doi: 10.1039/c2an35340g. Epub 2012 Jun 15.
8
Direct and mediated electrochemistry of peroxidase and its electrocatalysis on a variety of screen-printed carbon electrodes: amperometric hydrogen peroxide and phenols biosensor.过氧化物酶的直接和介导电化学及其在各种丝网印刷碳电极上的电催化作用：安培型过氧化氢和酚类生物传感器
Anal Bioanal Chem. 2015 Jan;407(2):439-46. doi: 10.1007/s00216-014-8282-x. Epub 2014 Nov 6.
9
Voltammetric determination of alkaline phosphatase and horseradish peroxidase activity using 3-indoxyl phosphate as substrate Application to enzyme immunoassay.
Talanta. 2004 Oct 8;64(2):452-7. doi: 10.1016/j.talanta.2004.03.003.
10
Disposable biosensors for determination of biogenic amines.一次性生物传感器用于测定生物胺。
Anal Chim Acta. 2010 Apr 14;665(1):26-31. doi: 10.1016/j.aca.2010.03.012. Epub 2010 Mar 12.
引用本文的文献
1
Development of quantitative detection methods for four Alzheimer's disease specific biomarker panels using electrochemical immunosensors based on enzyme immunoassay.基于酶联免疫测定的电化学免疫传感器用于四种阿尔茨海默病特异性生物标志物检测试剂盒的定量检测方法的开发。
Anal Sci. 2024 Oct;40(10):1809-1821. doi: 10.1007/s44211-024-00614-7. Epub 2024 Jun 17.
2
Carbon dots derived from expired drugs based ratiometric fluorescent sensor for horseradish peroxidase in fruits and vegetables and screening inhibitors.基于过期药物的碳点比率荧光传感器用于水果和蔬菜中辣根过氧化物酶的检测及抑制剂的筛选
Mikrochim Acta. 2024 Jan 22;191(2):109. doi: 10.1007/s00604-023-06160-9.
3
Polyaniline Entrapped Water-Dispersible 3MPA-ZnSe Quantum Dots and Their Application for the Development of an Enzymatic Electrochemical Nanobiosensor for the Detection of 17β-Estradiol, an Endocrine-Disrupting Compound.聚邻苯二胺包裹的水分散性 3MPA-ZnSe 量子点及其在开发用于检测内分泌干扰物 17β-雌二醇的酶电化学纳米生物传感器中的应用。
Appl Biochem Biotechnol. 2023 May;195(5):3425-3455. doi: 10.1007/s12010-022-04277-w. Epub 2023 Jan 4.
4
Voltammetric determination of the Escherichia coli DNA using a screen-printed carbon electrode modified with polyaniline and gold nanoparticles.基于聚苯胺和金纳米粒子修饰的丝网印刷碳电极的大肠杆菌 DNA 的伏安测定法。
Mikrochim Acta. 2018 Mar 12;185(4):217. doi: 10.1007/s00604-018-2749-y.
5
Design, Synthesis, and Characterization of Small-Molecule Reagents That Cooperatively Provide Dual Readouts for Triaging and, When Necessary, Quantifying Point-of-Need Enzyme Assays.用于即时检测酶分析的小分子试剂的设计、合成与表征，这些试剂可协同提供双重读数，用于分类，并在必要时进行定量。
J Org Chem. 2015 Nov 6;80(21):10437-45. doi: 10.1021/acs.joc.5b02013. Epub 2015 Oct 16.
本文引用的文献
1
Characterization of new fluorescent peroxidase substrates.新型荧光过氧化物酶底物的特性研究。
Talanta. 2003 Jun 13;60(2-3):345-54. doi: 10.1016/S0039-9140(03)00097-3.
2
Detection of ferritin in human serum with a MAP-H(2)O(2)-HRP voltammetric enzyme-linked immunoassay system.采用 MAP-H2O2-HRP 伏安酶联免疫分析系统检测人血清中的铁蛋白。
Talanta. 1999 Aug 23;50(1):95-101. doi: 10.1016/s0039-9140(99)00105-8.
3
Differential pulse voltammetric enzyme-linked immunoassay for the determination of Helicobacter pylori specific immunoglobulin G (IgG) antibody.差分脉冲伏安酶联免疫分析法测定幽门螺杆菌特异性免疫球蛋白G（IgG）抗体
Talanta. 1997 May;44(5):823-30. doi: 10.1016/s0039-9140(96)02120-0.
4
The effects of media properties on the horseradish peroxidase-catalyzed fluorogenic reaction.
Talanta. 1994 Dec;41(12):2049-54. doi: 10.1016/0039-9140(94)00177-4.
5
Detection of two different mRNAs in a single section by dual in situ hybridization: a comparison between colorimetric and fluorescent detection.通过双重原位杂交在单个切片中检测两种不同的mRNA：比色法和荧光检测之间的比较。
J Neurosci Methods. 2007 May 15;162(1-2):119-28. doi: 10.1016/j.jneumeth.2006.12.017. Epub 2007 Jan 12.
6
Production of anti-digoxigenin antibody HRP conjugate for PCR-ELISA DIG detection system.用于PCR-ELISA地高辛检测系统的抗地高辛抗体辣根过氧化物酶偶联物的制备。
J Immunoassay Immunochem. 2006;27(4):303-18. doi: 10.1080/15321810600861910.
7
Layer-by-layer hydroxymethyl ferrocene modified sensor for one-step flow/stop-flow injection amperometric immunoassay of alpha-fetoprotein.用于甲胎蛋白一步流动/停流注射安培免疫分析的逐层羟甲基二茂铁修饰传感器。
Biosens Bioelectron. 2007 Mar 15;22(8):1700-6. doi: 10.1016/j.bios.2006.07.034. Epub 2006 Sep 1.
8
Biosensor based on self-assembling acetylcholinesterase on carbon nanotubes for flow injection/amperometric detection of organophosphate pesticides and nerve agents.基于碳纳米管上自组装乙酰胆碱酯酶的生物传感器，用于流动注射/安培法检测有机磷农药和神经毒剂。
Anal Chem. 2006 Feb 1;78(3):835-43. doi: 10.1021/ac051559q.
9
Continuous-flow/stopped-flow system for enzyme immunoassay using a rotating bioreactor: determination of Chagas disease.
Biosens Bioelectron. 2005 Aug 15;21(2):313-21. doi: 10.1016/j.bios.2004.09.029. Epub 2004 Nov 26.
10
Amperometric detection in TMB/HRP-based assays.基于TMB/HRP检测法中的安培检测
Anal Bioanal Chem. 2005 May;382(2):297-302. doi: 10.1007/s00216-005-3084-9. Epub 2005 May 13.
Zotero 插件