Development of DNA aptamer-based sensor for electrochemical detection of C-reactive protein
Development of DNA aptamer-based sensor for electrochemical detection of C-reactive protein
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DOI：
10.1016/j.talanta.2018.06.035
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发表时间：
2018-11-01
期刊：
影响因子：
6.1
通讯作者：
Malinowska, Elzbieta
中科院分区：
文献类型：
--
作者：
Jarczewska, Marta;Rebis, Janusz;Malinowska, Elzbieta
C-反应蛋白（CRP）是心血管疾病的重要生物标志物，其检测应用了光学和电化学技术。本研究将DNA适体作为识别层用于CRP的检测。为此，选择了金表面上的适体固定方法，并优化了受体层的含量以确保与靶蛋白的有效结合。通过计时库仑法和原子力显微镜的应用，单层的质量进行了验证。使用硫醇化适体提供了最高密度和稳定性的层的形成。在亚甲基蓝氧化还原指示剂存在下进行的方波伏安法实验揭示了在1至100 pM的范围内适体传感器对CRP的线性响应。此外，与包括BSA和IgE的干扰蛋白相比，基于DNA适体的传感器显示出对C-反应蛋白的良好选择性。最后，使用所开发的适体传感器对血清样本中的CRP进行分析。
C-reactive protein (CRP) is a crucial biomarker of cardiovascular diseases and for its detection both optical and electrochemical techniques were applied. This study concerns the application of DNA aptamer as recognition layer for CRP detection. For that purpose aptamer immobilization method on gold surface was selected and the content of receptor layer was optimized to ensure an efficient binding to target protein. The quality of the monolayer was verified by the application of chronocoulometry and atomic force microscopy. Using thiolated aptamers provided the formation of layers of highest density and stability. The square-wave voltammetry experiments performed in the presence of methylene blue redox indicator revealed a linear response of aptasensor towards CRP in the range from 1 to 100 pM. Moreover, a DNA aptamer - based sensor showed good selectivity towards C-reactive protein in comparison to interfering proteins including BSA and IgE. Finally, the analysis of CRP in serum sample was conducted using the developed aptasensor.