Selection of broad-spectrum aptamer and its application in fabrication of aptasensor for detection of aminoglycoside antibiotics residues in milk
Selection of broad-spectrum aptamer and its application in fabrication of aptasensor for detection of aminoglycoside antibiotics residues in milk
复制标题
广谱适配体的筛选及其在检测牛奶中氨基糖苷类抗生素残留适配体传感器中的应用
DOI：
10.1016/j.snb.2021.130959
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发表时间：
2021-10
影响因子：
8.4
通讯作者：
Xia Sun
中科院分区：
文献类型：
--
作者：
Fengling Yue;He Li;Qianqian Kong;Jing Liu;Guangxian Wang;Falan Li;Qingqing Yang;Wei Chen;Xia Sun
据报道，食品中存在多种氨基糖苷类抗生素残留。将广谱核酸适体作为传感器的识别元件，对该类氨基酸的同时检测具有重要意义。本文利用氧化石墨烯（GO）-SELEX技术，以一组靶点为靶点，成功筛选出了具有高亲和性和特异性的广谱单链DNA适配子。进一步序列分析，观察到仅49 nt的截短适体（AAs 03-2）具有比79 nt的原始适体更高的亲和力。通过圆二色谱和分子对接模拟分析了适配体（AAs 03-2）的结合机制，发现适配体的折叠结构为AAs提供了结合口袋，并能识别AAs的保守结构。随后，进一步验证了适体（AAs 03-2）对具有有利结合能力的其他AAs的亲和力。最后，利用该核酸适体（AAs 03-2）构建了基于有序介孔碳修饰丝网印刷碳电极的电化学生物传感器，用于信号放大。该适体传感器实现了AAs的同时分析，检测限为2.47 nM。该方法成功应用于牛奶中氨基酸残留的检测，为抗生素多残留检测提供了新的方法。·通过GO-SELEX成功地选择了具有高亲和力的AAs类的广谱ssDNA适体。·适体的折叠结构为氨基酸提供了结合口袋，并可识别氨基酸的保守结构。·以所选适体为识别元件的适体传感器实现了牛奶中AA的同时检测。
The multi-residues of aminoglycoside antibiotics (AAs) have been reported present in a food product. Taking the broad-spectrum aptamer as recognition element of sensors for simultaneous detection of the class of AAs is of great significance. In this paper, broad-spectrum ssDNA aptamers for AAs with high affinity and specificity were successfully selected by graphene oxide (GO)-SELEX with a group of targets. Further sequence analysis, the truncated aptamer (AAs 03-2) of only 49 nt was observed with higher affinity than original aptamer of 79 nt. The binding mechanism analysis was performed by circular dichroism and molecular docking simulations, which showed the folding structure of aptamer (AAs 03-2) provided binding pockets for AAs and could recognize AAs conserved structure. Subsequently, the affinity of aptamer (AAs 03-2) for other AAs with favorable binding ability was further verified. Finally, the aptamer (AAs 03-2) was used to fabricate electrochemical biosensor based on screen-printed carbon electrode modified by ordered mesoporous carbon for signal amplification. The aptasensor realized simultaneous analysis of AAs with limit of detection of 2.47 nM. Moreover, the proposed aptasensor was successfully applied to evaluate the residual level of AAs in milk, which provides a new method for multi-residues detection of antibiotics. • The broad-spectrum ssDNA aptamers for the class of AAs with high affinity were successfully selected by GO-SELEX. • The folding structure of aptamer provided binding pockets for AAs and could recognize conserved structure of AAs. • The aptasensor with the selected aptamer as recognition element realized simultaneous detection of AAs in milk.
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DOI：
10.1016/j.snb.2021.130377
发表时间：
2021-10-15
期刊：
Sensors and actuators. B, Chemical
影响因子：
--
作者：
Amouzadeh Tabrizi M;Nazari L;Acedo P
通讯作者：
Acedo P
影响因子：
7.4
作者：
Derbyshire, Nicola;White, Simon J.;Bunka, David H. J.;Song, Lei;Stead, Sara;Tarbin, Jonathan;Sharman, Matthew;Zhou, Dejian;Stockley, Peter G.
通讯作者：
Stockley, Peter G.
影响因子：
5.7
作者：
Han, Xuyan;Zhang, Yuhong;Zhou, Nandi
通讯作者：
Zhou, Nandi
影响因子：
64.8
作者：
Parisien, Marc;Major, Francois
通讯作者：
Major, Francois
影响因子：
4.1
作者：
Dasenaki, Marilena E.;Michali, Christina S.;Thomaidis, Nikolaos S.
通讯作者：
Thomaidis, Nikolaos S.