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基于富含鸟嘌呤互补 DNA 序列用于信号放大的适体/金纳米颗粒/硫化钴纳米片生物传感器用于检测 17β-雌二醇

Aptamer/Au nanoparticles/cobalt sulfide nanosheets biosensor for 17β-estradiol detection using a guanine-rich complementary DNA sequence for signal amplification.

机构信息

College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China; State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.

College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.

出版信息

Biosens Bioelectron. 2015 May 15;67:184-91. doi: 10.1016/j.bios.2014.08.010. Epub 2014 Aug 15.

Abstract

We have developed a sensitive sensing platform for 17β-estradiol by combining the aptamer probe and hybridization reaction. In this assay, 2-dimensional cobalt sulfide nanosheet (CoS) was synthesized by a simple hydrothermal method with L-cysteine as sulfur donor. An electrochemical aptamer biosensor was constructed by assembling a thiol group tagged 17β-estradiol aptamer on CoS and gold nanoparticles (AuNPs) modified electrode. Methylene blue was applied as a tracer and a guanine-rich complementary DNA sequence was designed to bind with the unbound 17β-estradiol aptamer for signal amplification. The binding of guanine-rich DNA to the aptamer was inhibited when the aptamer captured 17β-estradiol. Using guanine-rich DNA in the assay greatly amplified the redox signal of methylene blue bound to the detection probe. The CoS/AuNPs film formed on the biosensor surface appeared to be a good conductor for accelerating the electron transfer. The method demonstrated a high sensitivity of detection with the dynamic concentration range spanning from 1.0×10(-9) to 1.0×10(-12) M and a detection limit of 7.0×10(-13) M. Besides, the fabricated biosensor exhibited good selectivity toward 17β-estradiol even when interferents were presented at 100-fold concentrations. Our attempt will extend the application of the CoS nanosheet and this signal amplification assay to biosensing areas.

摘要

我们结合适体探针和杂交反应开发了一种用于检测 17β-雌二醇的灵敏传感平台。在该测定法中,通过简单的水热法以 L-半胱氨酸作为硫供体合成了二维钴硫化物纳米片(CoS)。通过将巯基标记的 17β-雌二醇适体组装在 CoS 和金纳米粒子(AuNPs)修饰电极上,构建了电化学适体生物传感器。亚甲基蓝被用作示踪剂,设计了富含鸟嘌呤的互补 DNA 序列与未结合的 17β-雌二醇适体结合以进行信号放大。当适体捕获 17β-雌二醇时,富含鸟嘌呤的 DNA 与适体的结合受到抑制。在测定中使用富含鸟嘌呤的 DNA 极大地放大了与检测探针结合的亚甲基蓝的氧化还原信号。在生物传感器表面形成的 CoS/AuNPs 薄膜似乎是一种良好的导体,可加速电子转移。该方法的检测灵敏度高,动态浓度范围从 1.0×10(-9)到 1.0×10(-12) M,检测限为 7.0×10(-13) M。此外,即使在 100 倍浓度的干扰物存在下,所制备的生物传感器对 17β-雌二醇仍表现出良好的选择性。我们的尝试将扩展 CoS 纳米片的应用和这种信号放大测定法在生物传感领域的应用。

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