基于二硫化钼/纳米金和硫堇/纳米金信号放大的雌二醇电化学适配体传感器

doi:10.7503/cjcu20190199

摘要/Abstract

摘要：

构建了一种新型的基于二硫化钼/纳米金和硫堇/纳米金信号放大的检测17β-雌二醇的电化学适配体传感器. 利用巯基自组装技术将17β-雌二醇的适配体探针DNA固定在二硫化钼/纳米金修饰玻碳电极表面, 与末端带巯基的部分互补DNA链杂交, 将硫堇/纳米金电化学指示剂自组装在杂交后的双链DNA上, 制备了17β-雌二醇电化学适配体传感器. 二硫化钼/纳米金复合材料增加了电极的有效表面积和DNA探针的固定量. 纳米金作为信号物质载体负载硫堇, 实现了电化学指示剂的信号放大. 加入目标物17β-雌二醇后, 目标物与适配体DNA特异性结合, 导致互补DNA链脱落, 双链上结合的硫堇/纳米金电化学指示剂数量减少, 电化学信号降低. 实验结果表明, 在1.0×10 ^-14~5.0×10 ^-12 mol/L范围内17β-雌二醇浓度与峰电流的线性关系良好, 检出限为4.2×10 ^-15 mol/L(S/N=3). 该传感器可望用于其它环境激素类物质的检测.

关键词: 17β-雌二醇, 二硫化钼, 纳米金, 硫堇, 电化学适配体传感器

Abstract:

An ultrasensitive electrochemical aptasensor for the determination of 17β-estradiol(E2) was constructed based on signal-enhanced with molybdenum disulfide nanosheet(MoS₂)-gold nanoparticles(GNPs) and thionine-gold nanoparticle(THI/GNPs) conjugates as electrochemical indicator. The aptasensor was fabricated in sequence by self-assembling of thiolated aptamer DNA probes onto the surfaces of the modified electrodes, and then hybridized with thiolated complementary strand DNA(csDNA) to form double-stranded DNA(dsDNA) duplex. MoS₂-GNPs nanoparticles were used to increase the electrode surface area, enhance the electron transfer rate and amplify the sensor signal. GNPs as a biomarker load thionine, implements the electrochemical indicator signal amplification. E2 can be detected in the concentration range from 1.0×10 ^-14 mol/L to 5.0×10 ^-12 mol/L, and down to levels as low as 4.2×10 ^-15 mol/L. This study may offer a new approach with good analytical properties and potential applications.

Key words: 17β-Estradiol, Molybdenum disulfide nanosheets, Gold nanoparticles, Thionine, Electrochemical aptasensor

中图分类号:

O657.1

TrendMD:

贺彩梅,郑景伊,李晓霞. 基于二硫化钼/纳米金和硫堇/纳米金信号放大的雌二醇电化学适配体传感器. 高等学校化学学报, 2019, 40(10): 2090.

HE Caimei,ZHENG Jingyi,LI Xiaoxia. MoS₂-Gold Nanoparticles and Thionine-gold Nanoparticle Based Signal-enhanced Electrochemical Aptasensor for the Detection of 17β-Estradiol ^†. Chem. J. Chinese Universities, 2019, 40(10): 2090.

图/表 9

Scheme 1 Schematic illustration of the aptasensor preparation and the proposed mechanism for E2 detection

Fig.1 SEM images(A, B) and EDS(C, D) spectra of the MoS2 nanosheet(A, C) and MoS2-GNPs(B, D)

Fig.2 CV(A) and EIS(B) curves of interfaces at different modified electrodes in 5.0 mmol/L [Fe(CN)6]3-/4-/10 mmol/L KCl The scan rate of CV(A) was 50 mV/s, the frequency range of EIS(B) was from 100 kHz to 1.0 Hz and the biased potential was 0.21 V. The inset shows the equivalent circuit diagram applied to fit the impedance spectroscopy. a. GCE; b. GNPs-MoS2/GCE; c. ssDNA/GNPs-MoS2/GCE; d. dsDNA/GNPs-MoS2/GCE; e. GNPs/dsDNA/GNPs-MoS2/GCE.

Fig.3 CV response of the electrochemical apta-sensor interacted with different concentrations of E2 in 0.01 mol/L Tris-HCl solution(pH=7.4) cE2/(mol·L-1): a. 0; b. 1.0×10-13; c. 1.0×10-12.

Fig.4 Optimization of immobilization time of aptamer DNA(A), GNPs(B), and the binding time of E2(C)

Fig.5 DPV profiles of the aptasensor for the detection different concentrations of E2 and plot of current response vs. E2 concentration(inset) cE2/(mol·L-1): a. 0; b. 1.0×10-14; c. 5.0×10-14; d. 1.0×10-13; e. 5.0×10-13; f. 1.0×10-12 ; g. 5.0×10-12.

Table 1 Comparison of electrochemical aptasensor for the detection of E2*

Electrode	Technique	Linear range/ (mol·L^-1)	LOD/(mol·L^-1)	Sample	Ref.
Aptamer/poly(Py-co-PAA)/GCE	EIS	1.0×10^-15—1.0×10^-6	1.0×10^-15	Urine water	[14]
Aptamer/CuS/GNPs/glucose oxidase/GCE	DPV	5.0×10^-13—5.0×10^-9	6.0×10^-14	Urine	[15]
Aptamer /GNPs/WS₂/GCE	DPV	1.0×10^-11—1.0×10^-8	1.0×10^-12	Serum water	[16]
Aptamer/Au electrode	EIS	1.0×10^-11—1.0×10^-8	2.0×10^-12	Urine	[17]
Aptamer /GNPs/CoS/GCE	DPV	1.0×10^-12—1.0×10^-9	7.0×10^-13	Urine	[18]
Aptamer/dendritic Au/BDD	EIS	1.0×10^-14 —1.0×10^-9	5.0×10^-15	Water	[19]
Aptamer/gold electrode chip	SWV	1.0×10^-11—1.0×10^-9	1.0×10^-13		[20]
Aptamer/CdSe nanoparticles-modified TiO₂ nanotube arrays	PEC	5.0×10^-14—1.5×10^-11	3.3×10^-14	Water	[21]
Aptamer/ GNPs-MoS₂/GCE	DPV	1.0×10^-14—5.0×10^-12	4.2×10^-15	Water	This work

Fig.6 Histograms of response of the aptasensor interaction with E2 and E2 analogues cEZ=1.0×10-12 mol/L. a. E2; b. BPA; c. HQ; d. E3.

Table 2 Detection of E2 in tap water samples using the electrochemical aptasensor(n=3)

Sample	Added/(mol·L^-1)	Found/(mol·L^-1)	RSD(%)	Recovery(%)
1	1.00×10^-14	1.05×10^-14	3.7	105.0
2	5.00×10^-14	5.32×10^-14	4.5	106.4
3	1.00×10^-13	9.89×10^-14	4.2	98.9

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