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基于外切酶 III 辅助信号放大的无标记电化学磁适体传感器用于癌胚抗原的测定。

A label-free electrochemical magnetic aptasensor based on exonuclease III-assisted signal amplification for determination of carcinoembryonic antigen.

机构信息

School of Pharmacy, Nanjing Medical University, Nanjing, 211166, Jiangsu, China.

出版信息

Mikrochim Acta. 2020 Aug 8;187(9):492. doi: 10.1007/s00604-020-04457-7.

DOI:10.1007/s00604-020-04457-7
PMID:32770422
Abstract

A novel label-free and exonuclease III (Exo III)-assisted signal amplification electrochemical aptasensor was constructed for the determination of carcinoembryonic antigen (CEA) via magnetic field-induced self-assembly of magnetic biocomposites (FeO@Au NPs-S1-S2-S3). The magnetic biocomposites were acquired by modifying double-stranded DNA (S1-S2-S3) on the surface of FeO@Au nanoparticles (FeO@Au NPs). Among them, FeO@Au NPs were used as carriers for magnetic separation, thiolated single-stranded DNA (S1) provided signal sequence, CEA aptamer (S2) worked as a recognition element, and complementary strand (S3) was used to form double strands. In the presence of CEA, S2 bonded with CEA competitively; the exposed S1 could not be cleaved since Exo III was inactive against ssDNA. The G-quadruplex/hemin complexes finally formed with the existence of K, and the high electrochemical signal of G-quadruplex/hemin complexes was recorded by differential pulse voltammetry (DPV) at - 0.6 V. Conversely, in the absence of CEA, dsDNA was cleaved from the 3' blunt end by Exo III; the disappearance of G-rich sequence blocked the generation of the signal. This method exhibited good selectivity and sensitivity for the determination of CEA; the linear range was from 0.1 to 200 ng mL and the limit of detection was 0.4 pg mL. Graphical abstract.

摘要

一种新型的无标记和外切核酸酶 III(Exo III)辅助信号放大电化学适体传感器,通过磁场诱导磁性生物复合材料(FeO@Au NPs-S1-S2-S3)的自组装来测定癌胚抗原(CEA)。磁性生物复合材料是通过在 FeO@Au 纳米粒子(FeO@Au NPs)表面修饰双链 DNA(S1-S2-S3)获得的。其中,FeO@Au NPs 用作磁分离的载体,巯基化单链 DNA(S1)提供信号序列,CEA 适体(S2)作为识别元件,互补链(S3)用于形成双链。在存在 CEA 的情况下,S2 与 CEA 竞争结合;由于 Exo III 对 ssDNA 无活性,因此未被切割的 S1 无法被切割。在 K 的存在下最终形成 G-四链体/血红素复合物,并用差分脉冲伏安法(DPV)在-0.6 V 记录 G-四链体/血红素复合物的高电化学信号。相反,在没有 CEA 的情况下,Exo III 从 3'钝端切割 dsDNA;富含 G 的序列的消失阻止了信号的产生。该方法对 CEA 的测定表现出良好的选择性和灵敏度;线性范围为 0.1 至 200 ng mL,检测限为 0.4 pg mL。图表摘要。

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