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基于靶标触发适体信标和链霉亲和素-无机杂化复合材料的变构开关的无标记 IFN-γ 适体传感器。

A label-free IFN-γ aptasensor based on target-triggered allosteric switching of aptamer beacon and streptavidin-inorganic hybrid composites.

机构信息

College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou, 450001, PR China.

College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou, 450001, PR China.

出版信息

Anal Chim Acta. 2019 Dec 9;1087:29-35. doi: 10.1016/j.aca.2019.08.034. Epub 2019 Aug 17.

Abstract

A label-free electrochemical aptasensor was developed for the sensitive detection of interferon-gamma (IFN-γ). To do this, a diblock dual-aptamer allosteric hairpin (DDAH) was designed, followed by conjugation with gold nanoparticles (DDAH&AuNP). The presence of target destroyed the stable hairpin structure, and then the catalytic cleavage of DNAzymes removed the IFN-γ-binding molecules, triggering the allosteric switching from inactive hairpin to active streptavidin aptamer (A-DDAH&AuNP) in homogeneous system. Moreover, streptavidin-inorganic hybrid nanoflowers decorated with graphene composites (SFG) were synthesized and used as substrates to modify glassy carbon electrodes (SFG/GCE). SFG specifically bind to the A-DDAH&AuNP to realize high-efficient readout of signals. Under the optimal conditions and by using differential pulse stripping voltammetry (DPSV), the response peak currents increases linearly with the logarithm of the IFN-γ concentration in the range between 0.1 pg mL and 500 ng/mL. The detection limit is as low as 19 fg mL. The aptasensor also has excellent electrochemical performances, which exhibits broad application prospects in biometric analysis.

摘要

一种无标记电化学适体传感器被开发用于灵敏检测干扰素-γ(IFN-γ)。为此,设计了一种双嵌段双适体变构发夹(DDAH),随后与金纳米粒子(DDAH&AuNP)缀合。目标物的存在破坏了稳定的发夹结构,然后 DNA 酶的催化裂解去除了 IFN-γ 结合分子,触发了从无活性发夹到同质体系中活性链霉亲和素适体(A-DDAH&AuNP)的变构开关。此外,还合成了具有石墨烯复合材料修饰的链霉亲和素-无机杂化纳米花(SFG),并用作修饰玻碳电极(SFG/GCE)的基底。SFG 特异性结合 A-DDAH&AuNP 以实现信号的高效读出。在最佳条件下,通过差分脉冲溶出伏安法(DPSV),响应峰电流随 IFN-γ浓度的对数在 0.1 pg/mL 至 500 ng/mL 范围内呈线性增加。检测限低至 19 fg/mL。该适体传感器还具有优异的电化学性能,在生物计量分析中具有广阔的应用前景。

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