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基于发夹 DNA 的金纳米粒子增强 SPR 生物传感器,无非特异性吸附影响。

Au-NPs enhanced SPR biosensor based on hairpin DNA without the effect of nonspecific adsorption.

机构信息

College of Chemistry and Chemical Engineering, Graduate University of Chinese Academy of Sciences, Beijing 100049, PR China.

出版信息

Biosens Bioelectron. 2011 Jan 15;26(5):2473-7. doi: 10.1016/j.bios.2010.10.035. Epub 2010 Oct 30.

Abstract

Gold nanoparticles (Au-NPs) are usually used to amplify surface plasmon resonance (SPR) signals, however, the serious nonspecific adsorption has largely limited their practical applications. Here, we developed a novel Au-NPs enhanced biosensor without the effect of nonspecific adsorption: cutting Au-NPs off from the biosensor surface by RsaI endonuclease. In order to further improve the sensitivity, the probe DNA was designed specially. After the cleavage reaction, the residual probe DNA formed hairpin structure, which also resulted in a great change in SPR dip shift. Then, with the coaction of Au-NPs and conformation change of probe DNA, the SPR signal was amplified greatly. Using this method, we monitored the process of DNA cleavage in real-time and achieved a detection level of 5×10(-8) M. Moreover, the result of X-ray photoelectron spectroscopy (XPS) experiment further confirmed that large nonspecific adsorption existed. However, because SPR recorded a process in which the Au-NPs were cut off, the serious nonspecific adsorption did not affect the experimental result.

摘要

金纳米颗粒(Au-NPs)通常用于放大表面等离子体共振(SPR)信号,但严重的非特异性吸附在很大程度上限制了它们的实际应用。在这里,我们开发了一种新型的 Au-NPs 增强生物传感器,该传感器不受非特异性吸附的影响:通过 RsaI 内切酶将 Au-NPs 从生物传感器表面切割下来。为了进一步提高灵敏度,专门设计了探针 DNA。在切割反应后,残留的探针 DNA 形成发夹结构,这也导致 SPR 光密度位移发生了很大的变化。然后,在 Au-NPs 和探针 DNA 构象变化的共同作用下,SPR 信号被大大放大。使用这种方法,我们实时监测了 DNA 切割过程,实现了 5×10(-8) M 的检测水平。此外,X 射线光电子能谱(XPS)实验的结果进一步证实了存在大量的非特异性吸附。然而,由于 SPR 记录了 Au-NPs 被切割的过程,严重的非特异性吸附并不影响实验结果。

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