磁性印迹纳米粒子固定血红蛋白修饰磁性电极构建过氧化氢传感器

doi:10.13208/j.electrochem.180704

摘要/Abstract

摘要： 采用表面印迹技术，以磁性二氧化硅纳米粒子（Fe₃O₄@SiO₂ NPs）作为载体、血红蛋白（Hb）为模板分子、正硅酸乙酯（TEOS）为印迹聚合物单体，制备了Hb印迹Fe₃O₄@SiO₂的磁性印迹纳米粒子（MMIPs NPs）. MMIPs NPs具有磁性内核和血红蛋白印迹壳层的核壳结构，可以富集并固定Hb. 使用壳聚糖将MMIPs NPs固定于磁性电极表面，构建血红蛋白类酶生物传感器，研究了Hb对过氧化氢（H₂O₂）的催化活性. MMIPS NPS相比于磁性非印迹纳米粒子（MNIPS NPS），催化电流增加了14.3%. 采用磁性电极，MMIPS NPS、Hb和O₂的顺磁性使得该类酶生物传感器对H₂O₂的催化电流增加了60.0%. 血红蛋白类酶生物传感器电流响应与H₂O₂浓度在25 ~ 200 μmol·L^-1间呈线性关系，检出限为3 μmol·L^-1（S/N=3），表明该类酶传感器对H₂O₂具有良好的催化性能.

关键词: 血红蛋白, 类酶传感器, 过氧化氢, 磁性印迹纳米粒子, 磁性电极

Abstract: In this work, the surface-imprinted technique was used to prepare magnetic hemoglobin (Hb) imprinted nanoparticles, using Fe₃O₄@SiO₂ NPs as the carrier, Hb as the template molecule, and tetraethyl orthosilicate (TEOS) as the imprinted polymer monomer. The nanoparticles had a core-shell structure, with magnetic Fe₃O₄ NPs as the core and Hb imprinted polymers as the shell. Therefore, Hb could be concentrated and fixed on the surface of the magnetic imprinted nanoparticles (MMIPs NPs). Furthermore, MMIPs NPs were immobilized with chitosan (CS) on the surface of a magnetic electrode to constitute Hb enzyme-like biosensor to catalyze the reduction of hydrogen peroxide (H₂O₂). Compared with magnetic non-imprinted polymer nanoparticles (MNIPs NPs), the MMIPs NPs biosensor enhanced the response by 14.3%. Notably, an introduction of a magnetic field made the biosensor more sensitive owing to the paramagnetism of MNIPs NPs, Hb and O₂ molecules. The reduction current of H₂O₂ on Hb/MMIPs NPs modified magnetic glassy carbon electrode increased by 60.0%. Under the optimum condition, the linear detection range of H₂O₂ was 25 ~ 200 μmol·L^-1 with the detection limit of 3 μmol·L^-1 (S/N = 3), which showed that Hb enzyme-like biosensor had a good catalytic performance for H₂O₂.

Key words: hemoglobin, enzyme-like biosensor, hydrogen peroxide, magnetic imprinted nanoparticles, magnetic electrode

中图分类号:

O646.5

袁洋, 王佳新, 曹玉华. 磁性印迹纳米粒子固定血红蛋白修饰磁性电极构建过氧化氢传感器[J]. 电化学（中英文）, 2019, 25(6): 757-763.

YUAN Yang, WANG Jia-xin, CAO Yu-hua. Electrochemical Sensor Based on Magnetic Electrode Modified with Magnetic Molecularly Imprinted Nanoparticles Immobilized Hemoglobin for Determination of Hydrogen Peroxide[J]. Journal of Electrochemistry, 2019, 25(6): 757-763.

参考文献

[1] Lin V S, Lippert A R, Chang C J. Cell-trappable fluorescent probes for endogenous hydrogen sulfide signaling and imaging H₂O₂-dependent H₂S production[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110(18): 7131-7135.
[2] Miller E W, Chang C J. Fluorescent probes for nitric oxide and hydrogen peroxide in cell signaling[J]. Current Opinion in Chemical Biology, 2007, 11(6): 620-625.
[3] Tian Y C, Fan M, Qin Z X, et al. Hydrogen peroxide positively regulates brassinosteroid signaling through oxidation of the BRASSINAZOLE-RESISTANT1 transcription factor[J]. Nature Communications, 2018, 9(1): 1063.
[4] Wu P（吴萍）, Cai C X（蔡称心）. Horseradish peroxidase-attapulgite clay nanocomposites: Fabrication and application to sensing the extracellular H₂O₂ released from cells[J]. Journal of Electrochemistry（电化学）, 2014, 20(3): 260-265.
[5] Liu H Y, Weng L Y, Yang C. A review on nanomaterial-based electrochemical sensors for H₂O₂, H2S and NO inside cells or released by cells[J]. Microchimica Acta, 2017, 184(5): 1267-1283.
[6] Yagati A K, Choi J. Protein based electrochemical biosensors for H₂O₂ detection towards clinical diagnostics[J]. Electroanalysis, 2014, 26(6): 1259-1276.
[7] Zhang S Y(张思宇), Wang H J(王会娟), Li S F(李书芳), et al. Carbon composite Fe₃O₄ nanoparticles based electrochemical sensor for hydrogen peroxide detection[J]. Journal of Electrochemistry(电化学), 2018, 24(3): 279-284.
[8] Zhang Q(张倩), Fu S Y(付时雨), Li H L(李海龙), et al. A rapid method for the determination of hydrogen peroxide concentration[J]. Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2104, 34(3): 767-770.
[9] Raja S, Ramesh V, Thivaharan V. Green biosynthesis of silver nanoparticles using Calliandra haematocephala, leaf extract, their antibacterial activity and hydrogen peroxide sensing capability[J]. Arabian Journal of Chemistry, 2017, 10(2): 253-261.
[10] Ding J, Zhong Q, Zhang S L, et al. Simultaneous removal of NO_X, and SO₂, from coal-fired flue gas by catalytic oxidation-removal process with H2O2[J]. Chemical Engineering Journal, 2014, 243(5): 176-182.
[11] Motaghed R M, Ge L, Jiang H, et al. A facile photoelectrochemical sensor for high sensitive ROS and AA detection based on graphitic carbon nitride nanosheets[J]. Biosensors & Bioelectronics, 2018, 107: 54-61.
[12] Qu P(瞿鹏), Li B X(李保新), Zhang Z J(章竹君). Plant tissue-based chemiluminescence flow biosensor for hydrogen peroxide determination in water samples[J]. Analytical Chemistry(分析化学), 2003, 31(10): 1240-1243.
[13] Jo E J, Mun H, Kim S J, et al. Detection of ochratoxin A (OTA) in coffee using chemiluminescence resonance energy transfer (CRET) aptasensor[J]. Food Chemistry, 2016, 194: 1102-1107.
[14] Zangheri M, Cevenini L, Anfossi L, et al. A simple and compact smartphone accessory for quantitative chemiluminescence-based lateral flow immunoassay for salivary cortisol detection[J]. Biosensors & Bioelectronics, 2015, 64: 63-68.
[15] Zhang L S, Wong G T F. Optimal conditions and sample storage for the determination of H₂O₂ in marine waters by the scopoletin-horseradish peroxidase fluorometric method[J]. Talanta, 1999, 48(5): 1031-1038.
[16] Zhang C, Wang X R, Hou M, et al. Immobilization on metal-organic framework engenders high sensitivity for enzymatic electrochemical detection[J]. ACS Applied Materials & Interfaces, 2017, 9(16): 13831-13836.
[17] Li L M, Du Z F, Liu S A, et al. A novel nonenzymatic hydrogen peroxide sensor based on MnO₂/graphene oxide nanocomposite[J]. Talanta, 2010, 82(5): 1637-1641.
[18] He Y P, Sheng Q L, Zheng J B, et al. Magnetite-graphene for the direct electrochemistry of hemoglobin and its biosensing application[J]. Electrochimica Acta, 2011, 56(5): 2471-2476.
[19] Sun B H, Ni X J, Cao Y H, et al. Electrochemical sensor based on magnetic molecularly imprinted nanoparticles modified magnetic electrode for determination of Hb[J]. Biosensors & Bioelectronics, 2017, 91: 354-358.
[20] Nagababu E, Rifkind J M. Reaction of hydrogen peroxide with ferrylhemoglobin: superoxide production and heme degradation[J]. Biochemistry, 2000, 39(40): 12503-12511.
[21] Sun J Y, Huang K J, Zhao S F, et al. Direct electrochemistry and electrocatalysis of hemoglobin on chitosan-room temperature ionic liquid-TiO₂-graphene nanocomposite film modified electrode[J]. Bioelectrochemistry, 2011, 82(2): 125-130.
[22] Gautam V, Singh K P, Yadav V L. Polyaniline/multiwall carbon nanotubes/starch nanocomposite material and hemoglobin modified carbon paste electrode for hydrogen peroxide and glucose biosensing[J]. International Journal of Biological Macromolecules, 2018, 111: 1124-1132.
[23] Zhao H Y, Zheng W, Meng Z X, et al. Bioelectrochemistry of hemoglobin immobilized on a sodium alginate-multiwall carbon nanotubes composite film[J]. Biosensors & Bioelectronics, 2009, 24(8): 2352-2357.
[24] Jian F F, Qiao Y B, Zhuang R R. Direct electrochemistry of hemoglobin in TATP film: Application in biological sensor[J]. Sensors & Actuators B: Chemical, 2007, 124(2): 413-420.
[25] Feng J J, Xu J J, Chen H Y. Synergistic effect of zirconium phosphate and Au nanoparticles on direct electron transfer of hemoglobin on glassy carbon electrode[J]. Journal of Electroanalytical Chemistry, 2005, 585(1): 44-50.