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多聚腺嘌呤介导的球形核酸用于界面识别卡那霉素。

Poly-adenine-mediated spherical nucleic acids for interfacial recognition of kanamycin.

机构信息

Shanghai Engineering Research Center for Food Rapid Detection, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, China.

出版信息

Mikrochim Acta. 2022 Mar 22;189(4):151. doi: 10.1007/s00604-022-05235-3.

Abstract

Kanamycin fluorescence aptasensors were created using a series of di-block oligonucleotide modified gold nanoparticles with various lengths of poly-adenine. In the presence of kanamycin, the double strand structure of the aptamer-reporter strand complex is disrupted, and the dye-labelled reporter strand detaches from the surface of gold nanoparticles, resulting in fluorescence recovery (Ex/Em = 485/520 nm). By adjusting the number of consecutive adenines, the programable aptamer density can be implemented on the gold nanoparticle surface, and the conformation of nucleic acid changed from lying-down to up-right. The apparent binding constant, binding kinetics, and limit of detection of the prepared aptasensors were carefully examined to explore the influence of surface density. Under the optimum condition, the aptasensor had a tenfold lower limit of detection than the thiolated aptamer modified one, as low as 23.6 nM, when a di-block oligonucleotide with twenty consecutive adenines tailed. In addition, satisfactory recoveries ranging from 96.33 to 99.47% were achieved in spiked milk samples with relative standard deviation of 1.2-6.9% (n = 3). This surface density regulation strategy holds great promise in other aptamer-based interfacial recognition and sensing. Schematic presentation of di-block oligonucleotide modified gold nanoparticle with different surface densities and its kanamycin sensing application.

摘要

基于聚腺苷酸修饰的一系列双嵌段寡核苷酸金纳米粒子,制备了卡那霉素荧光适体传感器。在卡那霉素存在的情况下,适体-报告分子链复合物的双链结构被破坏,染料标记的报告分子链从金纳米粒子表面脱离,导致荧光恢复(Ex/Em=485/520nm)。通过调整连续腺嘌呤的数量,可以在金纳米粒子表面实现可编程适体密度,并且核酸的构象从躺卧变为直立。仔细研究了制备的适体传感器的表观结合常数、结合动力学和检测限,以探讨表面密度的影响。在最佳条件下,当使用带有二十个连续腺嘌呤尾巴的双嵌段寡核苷酸时,该适体传感器的检测限比巯基化适体修饰的传感器低十倍,低至 23.6nM。此外,在添加牛奶样品中,回收率在 96.33%至 99.47%之间,相对标准偏差为 1.2-6.9%(n=3)。这种表面密度调节策略在其他基于适体的界面识别和传感中具有广阔的应用前景。不同表面密度的双嵌段寡核苷酸修饰金纳米粒子及其卡那霉素传感应用的示意图。

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