识别牛血红蛋白的分子印迹电化学传感器的制备与研究
Fabrication and Study of Molecularly Imprinted Electrochemical Sensor for Selective Recognition of Bovine Hemoglobin
摘 要
采用电化学聚合物法制备了一种用于识别牛血红蛋白(BHb)的分子印迹电化学传感器。玻碳电极表面经电化学法沉积一层纳米金,接着自组装一层L-半胱氨酸,对修饰电极表面功能化,引入氨基和羧基,增加蛋白质印迹识别位点,以BHb为模板,采用电化学诱导的丙烯酰胺氧化还原聚合反应形成分子印迹聚合物层,模板蛋白分子经洗脱形成分子印迹电化学传感器。其产生的电化学信号的变化与BHb的质量浓度在1.0~1.0×103mg·L-1范围有直接的关系,检出限(3σ)达0.5 mg·L-1。这种新型传感器具有较高灵敏度,可用于目标蛋白的识别。
电化学传感器
牛血红蛋白
纳米金
L-半胱氨酸
Molecular imprinting
Electrochemical sensor
BHb
Gold nanoparticles
L-cysteine
Abstract
A molecularly imprinted electrochemical sensor (MIECS) was prepared by electrochemical polymerization for recognition of bovine hemoglobin (BHb). Gold nanoparticles were electrodeposited on the surface of glassy carbon electrode, and then a layer of L-cysteine was self-assembled on the Au nanoparticles to functionize the surface of the modified electrode, and to introduce amino and carboxyl radicals to increase imprinted sites for recognition of protein. The molecularly imprinted polymer layer was synthesized by electrochemically induced redox polymerization of acrylamide using BHb as template, and the MIECS was formed after removing the template protein molecules by washing. The MIECS showed good response between the electrochemical signs and the mass concentration of BHb in the range of 1.0-1.0×103mg·L-1 with detection limit (3σ) of 0.5 mg·L-1. The proposed sensor exhibited to have high sensitivity, and could be used to identify the target protein.
中图分类号 O657.1 DOI 10.11973/lhjy-hx201512001
所属栏目 试验与研究
基金项目 国家自然科学基金资助项目(21405075);福建省自 然科学基金(2013J01052);福建省教育厅中青年教师教育科研项 目(JA14248;JA14257)
收稿日期 2014/12/11
修改稿日期
网络出版日期
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联系人作者李艳霞(yxli09@163.com)
备注李艳霞(1980-),女,江西九江人,讲师,博士,主要 研究方向为生物传感器。
引用该论文: LI Yan-xia,CHEN Yi-ting,HUANG Lu,WANG Xiu-ping. Fabrication and Study of Molecularly Imprinted Electrochemical Sensor for Selective Recognition of Bovine Hemoglobin[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2015, 51(12): 1633～1638
李艳霞,陈毅挺,黄露,王秀平. 识别牛血红蛋白的分子印迹电化学传感器的制备与研究[J]. 理化检验－化学分册, 2015, 51(12): 1633～1638
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【2】CHEN Hong-jun, ZHANG Zao-hui, CAI Rong, et al. Molecularly imprinted electrochemical sensor based on nickel nanoparticles-graphene nanocomposites modified electrode for determination of tetrabromobisphenol A［J］. Electrochimica Acta, 2014,117:385-392.
【3】张进,姚桃花,王燕青.分子印迹电化学传感器研究进展［J］.理化检验-化学分册, 2013,49(8):1017-1023.
【4】YOLA M L, EREN T, ATAR N. Molecularly imprinted electrochemical biosensor based on Fe@Au nanoparticles involved in 2-aminoethanethiol functionalized multi-walled carbon nanotubes for sensitive determination of cefexime in human plasma［J］. Biosensors and Bioelectronics, 2014,60:277-285.
【5】REDDY S M, SETTE G, PHAN Q. Electrochemical probing of selective haemoglobin binding in hydrogel-based molecularly imprinted polymers［J］. Electrochimica Acta, 2011,56(25):9203-9208.
【6】WHITCOMBE M J, CHIANELLA I, LARCOMBE L, et al. The rational development of molecularly imprinted polymer-based sensors for protein detection［J］. Chemical Society Reviews, 2011,40(3):1547-1571.
【7】LI Jian-ping, SHAO Yi-juan, YIN Wei-ling, et al. A strategy for improving the sensitivity of molecularly imprinted electrochemical sensors based on catalytic copper deposition［J］. Analytica Chimica Acta, 2014,817:17-22.
【8】SCHELLER F W, BISTOLAS N, LIU S, et al. Thirty years of haemoglobin electrochemistry［J］. Advances in Colloid and Interface Science, 2005,116(1/3):111-120.
【9】李琳,何锡文,陈朗星,等,表面印迹制备牛血红蛋白分子印迹聚苯乙烯磁性纳米球［J］.中国科学:B辑, 2009(9):989-989.
【10】GAO Rui-xia, MU Xin-ru, HAO Yi, et al. Combination of surface imprinting and immobilized template techniques for preparation of core-shell molecularly imprinted polymers based on directly amino-modified Fe3O4 nanoparticles for specific recognition of bovine hemoglobin［J］. Journal of Materials Chemistry B, 2014,2(12):1733-1741.
【11】WU Shou-guo, TAN Wen-guang, XU Hai-hong. Protein molecularly imprinted polyacrylamide membrane: for hemoglobin sensing［J］. Analyst, 2010,135(10):2523-2527.
【12】KAN Xian-wen, XING Zong-lan, ZHU An-hong, et al. Molecularly imprinted polymers based electrochemical sensor for bovine hemoglobin recognition［J］. Sensors and Actuators B: Chemical, 2012,168:395-401.
【13】LI Lu, YANG Ling-ling, XING Zong-lan, et al. Surface molecularly imprinted polymers-based electrochemical sensor for bovine hemoglobin recognition［J］. Analyst, 2013,138(22):6962-6968.
【14】NOBREGA M M, MARTINS V L, TORRESI R M, et al. One-step synthesis, characterization, and properties of emeraldine salt nanofibers containing gold nanoparticles［J］. Journal of Physical Chemistry C, 2014,118(8):4267-4274.
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【16】SONG Wei, LI Hui, LIANG Hua, et al. Disposable electrochemical aptasensor array by using in situ DNA hybridization inducing silver nanoparticles aggregate for signal amplification［J］. Analytical Chemistry, 2014,86(5):2775-2783.
【17】LI Yan-xia, LI Yu-jun, HONG Mei, et al. Highly sensitive protein molecularly imprinted electro-chemical sensor based on gold microdendrites electrode and prussian blue mediatedamplification［J］. Biosensors and Bioelectronics, 2013,42:612-617.
【18】MENAKER A, SYRITSKI V, REUT J, et al. Electrosynthesized surface-imprinted conducting polymer microrods for selective protein recognition［J］. Advanced Materials, 2009,21(22):2271-2275.
【19】VISWANATHAN S, RANI C, RIBEIRO S, et al. Molecular imprinted nanoelectrodes for ultra sensitive detection of ovariancancer marker［J］. Biosensors and Bioelectronics, 2012,33(1):179-183.
【20】LIN Zi-an, SUN Li-xiang, LIU Wei, et al. Synthesis of boronic acid-functionalized molecularly imprinted silica nanoparticles for glycoprotein recognition and enrichment［J］. Journal of Materials Chemistry B, 2014,2(6):637-643.
【21】ZHANG Wei, HE Xi-wen, CHEN Yang, et al. Molecularly imprinted polymer anchored on the surface of denatured bovine serum albumin modified CdTe quantum dots as fluorescent artificial receptor for recognition of target protein［J］. Biosensors and Bioelectronics, 2012,31(1):84-89.
【22】WANG Zong-hua, LI Feng, XIA Jian-fei, et al. An ionic liquid-modified graphene based molecular imprinting electrochemical sensor for sensitive detection of bovine hemoglobin［J］. Biosensors and Bioelectronics, 2014,61:391-396.
【23】KOMAROVA E, ALDISSI M, BOGOMOLOVA A. Design of molecularly imprinted conducting polymer protein-sensing films via substrate-dopant binding［J］. Analyst, 2015,140(4):1099-1106.
【24】HE Hong-yan, FU Guo-qi, WANG Yan, et al. Imprinting of protein over silica nanoparticles via surface graft copolymerization using low monomer concentration［J］. Biosensors and Bioelectronics, 2010,26(2):760-765.
【25】FANG Guo-zhen, ZHAI Xiao-rui, DENG Qi-liang, et al. Preparation and evaluation of lysozyme molecularly imprinted polymer film on the surface of multi-wall carbon nanotubes［J］. Current Organic Chemistry, 2012,16(12):1461-1467.
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