[1]孟凡飞,刘飞,刘畅,等.单壁碳纳米角+空心纳米铂链复合物的类夹心免疫新型穿孔素适配体电化学传感器的构建[J].第三军医大学学报,2018,40(01):76-81.
 MENG Fanfei,LIU Fei,LIU Chang,et al.Construction of aptamer electrochemical sensor based on SWCNHs@HPtCs complexes for perforin detection[J].J Third Mil Med Univ,2018,40(01):76-81.
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单壁碳纳米角+空心纳米铂链复合物的类夹心免疫新型穿孔素适配体电化学传感器的构建(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第01期
页码:
76-81
栏目:
基础医学
出版日期:
2018-01-15

文章信息/Info

Title:
Construction of aptamer electrochemical sensor based on SWCNHs@HPtCs complexes for perforin detection
作者:
孟凡飞刘飞刘畅蒋栋能蒲晓允
陆军军医大学新桥医院检验科
Author(s):
MENG Fanfei LIU Fei LIU Chang JIANG Dongneng PU Xiaoyun

Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China

关键词:
穿孔素电化学传感器单壁碳纳米角纳米铂二茂铁
Keywords:
perforin electrochemical sensors single-walled carbon nanohorns platinum ferrocene
分类号:
R313; R341; R392-33
文献标志码:
A
摘要:

目的    构建一种基于单壁碳纳米角(singlewalled carbon nanohorns,SWCNHs) 与空心纳米铂链(hollow Pt chains,HPtCs)复合物(SWCNHs@HPtCs)的电化学适配体传感器。方法     将单壁碳纳米管(singlewall carbon nanotubes,SWCNTs)溶液滴加在洁净的玻碳电极上(glassy carbon electrode, GCE),使之在电极表面形成一层膜,通过循环伏安法(cyclic voltammetry,CV)在电极表面电沉积纳米金(Au nanoparticles,AuNPs),得到AuNPs/ SWCNTs/GCE。进一步通过金硫键将穿孔素适配体固载在电极表面。制备SWCNHs@HPtCs复合物,固载穿孔素抗体和氨基二茂铁(aminoferrocene, AFc),当目标物穿孔素存在时,可与适配体和抗体形成夹心结构,使标记有AFc的SWCNHs@HPtCs复合物连接到电极表面,通过示差脉冲伏安法(differential pulse voltammetry,DPV)检测AFc还原峰电流值变化,实现对穿孔素的定量检测。结果     成功制备了SWCNHs@HPtCs复合物。反应底液的最佳pH值为6.5,复合物最佳孵育时间为60 min。在优化实验条件下,连续扫描50圈,峰电流值下降6.3%。穿孔素浓度在1~20 nmol/L范围内与DPV峰电流值呈良好的线性关系,检出限为0.56 nmol/L。结论    该电化学传感器具有电流响应快,灵敏度高,稳定性好,选择性好的特点,可应用于穿孔素的检测。
 

Abstract:

Objective     To construct an aptamer electrochemical sensor based on singlewalled carbon nanohorns (SWCNHs) and hollow Pt chains (HPtCs) complex (SWCNHs@HPtCs). Methods    A solution containing single-wall carbon nanotubes (SWCNTs) was added on a clean glassy carbon electrode (GCE) to form a film on the surface of the electrode. A film of nano-gold was electrodeposited on the surface of the electrode by cyclic voltammetry (CV) to form AuNPs/SWCNTs/GCE. The SWCNHs@HPtCs complex was prepared by immobilizing perforin aptamers on the electrode surface and labeling aminoferrocene (AFc) as a signal carrier. When the target existed, the aptamers, perforin and antibodies formed a sandwich structure, which would connect AFc labeled HPtCs@SWCNHs complex to the electrode surface. And then detection of differential pulse voltammetry (DPV) would display the changes of AFc reduction peak current to achieve the identification of perforin and quantitative detection. Results     The SWCNHs@HPtCs complex was prepared successfully. The optimal pH value of the detection solution was 6.5 and the optimal incubation time of the complex was 60 min. Under the optimal experimental conditions, the current was decreased by 6.3% after 50 cycles of consecutive scanning. There was a good linear relationship between the concentration of perforin in the range of 1 to 20 nmol/L and the peak current of the DPV. The detection limit was 0.56 nmol/L. Conclusion     Our prepared electrochemical sensor has the characteristics of fast current response, high sensitivity, good stability and good selectivity, and can be applied to the detection of perforin.

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更新日期/Last Update: 2018-01-12