自催化型石墨烯-金纳米探针的制备及用于非小细胞肺癌CYFRA21-1电化学检测

引用本文: 李瑞怡, 储红霞, 余敏仪, 李姝晗, 李在均. 自催化型石墨烯-金纳米探针的制备及用于非小细胞肺癌CYFRA21-1电化学检测[J]. 分析化学, 2020, 48(7): 871-880. doi: 10.19756/j.issn.0253-3820.191453 shu

Citation: LI Rui-Yi, CHU Hong-Xia, YU Min-Yi, LI Shu-Han, LI Zai-Jun. Synthesis of Self-catalytic Graphene-Gold Nanoprobe and Its Application in Electrochemical Detection of CYFRA21-1 in Non-small Cell Lung Cancer[J]. Chinese Journal of Analytical Chemistry, 2020, 48(7): 871-880. doi: 10.19756/j.issn.0253-3820.191453 shu

自催化型石墨烯-金纳米探针的制备及用于非小细胞肺癌CYFRA21-1电化学检测

1.
江南大学药学院, 无锡 214122;
2.
江南大学化学与材料工程学院, 无锡 214122
基金项目:
本文系国家重点研发计划项目（No.2018YFC1603001）和国家自然科学基金项目（No.21576115）资助

摘要: 早期诊断对提高肺癌患者生活质量和延长生存期至关重要。本研究通过热解柠檬酸与谷氨酰胺的混合物得到谷氨酰胺功能化石墨烯量子点（Gln-GQD），所制备的Gln-GQD与HAuCI₄反应形成Gln-GQD/Au复合物。Gln-GQD作为还原剂将Au³⁺转化为Au⁰，最终形成Au纳米晶体。Gln-GQD还作为稳定剂被固定于Au纳米晶体的表面，从而实现Gln-GQD与Au的杂交。扫描电镜、透射电镜、X-射线衍射和红外光谱分析表明，Gln-GQD/Au平均粒径为（31.2±0.15） nm，具有立方金的晶体结构，Au和N元素均匀分布于球形粒子表面，含有丰富的OH、-NH、-COOH等功能基团。将发夹DNA 2（H2）通过Au-S键连接到Au纳米粒子表面，再利用EDC/NHS活化实现Gln-GQD的-COOH与硫堇（Thi）的氨基缩合，得到H2-Gln-GQD/Au-Thi氧化还原探针，基于此构建了无酶放大电化学传感平台。在CYFRA21-1存在下，此传感平台的H2可与预先修饰在金电极表面的发夹DNA 1（H1）杂交，释放出一个CYFRA21-1分子，所释放的CYFRA21-1可直接用于下一次靶DNA循环。通过靶诱导的DNA自组装链式反应，一个CYFRA21-1靶分子可将多个氧化还原探针固定在金电极表面，从而产生显著的电化学信号放大效应。H2与H1的杂交实现对靶DNA的特异性响应，Thi在电极表面发生可逆性氧化还原反应，对靶DNA的响应产生电化学信号，而Gln-GQD/Au原位催化Thi的氧化还原反应将进一步放大检测信号。CYFRA21-1浓度在2~100000 fmol/L之间，其差分脉冲伏安峰电流随CYFRA21-1浓度增加而线性增大。本方法检出限为0.67 fmol/L（S/N=3），灵敏度明显优于文献报道的方法。本方法成功用于人血清中CYFRA21-1的电化学检测。

关键词:

English

Synthesis of Self-catalytic Graphene-Gold Nanoprobe and Its Application in Electrochemical Detection of CYFRA21-1 in Non-small Cell Lung Cancer

1.
School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, China;
2.
School of Chemical and Materials Engineering, Jiangnan University, Wuxi 214122, China
Received Date: 28 July 2019
Revised Date: 09 May 2020
Fund Project:
This work was supported by the National Key Research and Development Program of China (No. 2018YFC1603001) and the National Natural Science Foundation of China (No. 21576115).

Abstract: Early diagnosis is very important for improving the quality of life and prolonging the survival time of lung cancer patients. In this work, glutamine-functionalized graphene quantum dot (Glu-GQD) was prepared by pyrolysis of the mixture of citric acid and glutamine at 180℃ for 3 h. The resulting Gln-GQD was reacted with HAuCl₄ to produce Gln-GQD/Au hybrid. On one hand, the Gln-GQD as the reductant converted Au³⁺ into Au⁰, finally leading to the formation of Au nanocrystals. On the other hand, the Gln-GQD as the stabilizer was immobilized on the surface of Au nanocrystals to achieve the hybridization of Gln-GQD and Au nanoparticle. The results of scanning electron microscopy, transmission electron microscopy, X-ray diffraction and infrared spectroscopy analysis revealed that the Gln-GQD/Au offerred the crystal structure of cubic gold, in which the gold and nitrogen elements were uniformly distributed on the surface of spherical Au nanoparticles, with the average particle size of (31.2±0.15) nm and the rich of functional groups such as -OH, -NH- and -COOH. To obtain H2-Gln-GQD/Au-Thi redox probe, one hairpin DNA 2 (H2) was connected to the surface of Au nanoparticles via the Au-S bond, and then the condensation of -COOH in the Gln-GQD with -NH₂ in the thionine (Thi) by EDC/NHS activation was achieved. Further, the redox probe was used to construct electrochemical sensing platform coupled with the nonenzymatic amplification strategy. In the presence of CYFRA21-1, the H2 in the redox probe was hybridized with the hairpin DNA 1 (H1) modified on the surface of gold electrode to release one CYFRA21-1, which could be directly used in the next target DNA cycle. By the target-induced DNA self-assembly reaction, one CYFRA21-1 molecule transferred multiple redox probes to the surface of gold electrodes, resulting in a significant electrochemical signal amplification. In the detection of CYFRA21-1, the hybridization of H2 with H1 achieved the specific response to target DNA. Thi underwent reversible redox reaction on the electrode surface and produced the electrochemical signal in the response to target DNA. The Gln-GQD/Au in situ catalytzed the oxidation-reduction reaction of Thi. This would further amplify the detection signal. When the concentrations of CYFRA21-1 were in the range of 2-100000 fmol/L, the differential pulse voltammetric peak current increased linearly with the increase of CYFRA21-1 concentration. The detection limit was found to be 0.67 fmol/L (S/N=3). The sensitivity was much better than that of reported electrochemical sensors for CYFRA21-1. This method was successfully applied to the electrochemical detection of CYFRA21-1 in human serum samples.

Key words:

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

自催化型石墨烯-金纳米探针的制备及用于非小细胞肺癌CYFRA21-1电化学检测

English

Synthesis of Self-catalytic Graphene-Gold Nanoprobe and Its Application in Electrochemical Detection of CYFRA21-1 in Non-small Cell Lung Cancer

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

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

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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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

自催化型石墨烯-金纳米探针的制备及用于非小细胞肺癌CYFRA21-1电化学检测

English

Synthesis of Self-catalytic Graphene-Gold Nanoprobe and Its Application in Electrochemical Detection of CYFRA21-1 in Non-small Cell Lung Cancer

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

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

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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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs