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适体在金属有机框架上的多种吸附特性用于核酸检测

Multiple adsorption properties of aptamers on metal-organic frameworks for nucleic acid assay.

作者信息

Xie Xiaoyu, Ke Ruifang, Cheng Chen, Wang Yi-Hui, Song Zhen, Zhang Chang-Dong, Wang Huai-Song

机构信息

Key Laboratory of Drug Quality Control and Pharmacovigilance (Ministry of Education), China Pharmaceutical University, Nanjing, 210009, China; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an, 710061, China.

Key Laboratory of Drug Quality Control and Pharmacovigilance (Ministry of Education), China Pharmaceutical University, Nanjing, 210009, China; Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing, 210009, China.

出版信息

Biosens Bioelectron. 2021 Mar 15;176:112896. doi: 10.1016/j.bios.2020.112896. Epub 2020 Dec 15.

Abstract

Enrichment and detection of circulating free nucleic acids in biological samples have gained great attention for disease diagnosis or prognostic evaluation. Nanoscale metal-organic frameworks (NMOFs) have been used for aptamer-based nucleic acid sensing. In this work, different NMOFs, including ZIF-8, MIL-88, MIL-100, MIL-101, as well as Eu-TDA and Tb-TDA [prepared by the coordination of 2,2'-thiodiacetic acid (TDA) and Eu or Tb], were investigated in nucleic acid sensing by employing their aptamer adsorption ability and fluorescence quenching capacity for the labeled dyes. Two types of dye aptamer, FAM-labeled aptamer (FAM-Ap) and TexasRedaptamer (TexasRed-Ap) were designed, and their adsorption properties on NMOFs-were compared. It was found that the TexasRed-Ap can be well used for nucleic acid (miR-21) extraction and sensing by linking with a pH-responsive nucleotide chain (TexasRed-Ap-pH) or with an additional random chain ssDNA-1' (TexasRed-Ap-a). After interacted with the target miR-21 in biosamples, the TexasRed-dsDNA + NMOFs composites can be collected, and the formed TexasRed-dsDNA can be released by changing pH value or addition of ssDNA-1, which is matched with ssDNA-1'. A linear relationship from 0.1 to 200 pM for miR-21 detection was obtained. The results show that the NMOFs can be used as promising platforms for nucleic acid extraction and fluorescent sensing.

摘要

生物样品中循环游离核酸的富集和检测在疾病诊断或预后评估方面备受关注。纳米级金属有机框架(NMOF)已被用于基于适配体的核酸传感。在本工作中,通过利用其适配体吸附能力和对标记染料的荧光猝灭能力,研究了不同的NMOF,包括ZIF-8、MIL-88、MIL-100、MIL-101,以及Eu-TDA和Tb-TDA [由2,2'-硫代二乙酸(TDA)与Eu或Tb配位制备] 在核酸传感中的应用。设计了两种类型的染料适配体,FAM标记的适配体(FAM-Ap)和TexasRed适配体(TexasRed-Ap),并比较了它们在NMOF上的吸附特性。结果发现,TexasRed-Ap通过与pH响应核苷酸链(TexasRed-Ap-pH)或额外的随机链ssDNA-1'(TexasRed-Ap-a)连接,可很好地用于核酸(miR-21)的提取和传感。在与生物样品中的目标miR-21相互作用后,可收集TexasRed-dsDNA + NMOF复合材料,通过改变pH值或添加与ssDNA-1'匹配的ssDNA-1,可释放形成的TexasRed-dsDNA。获得了miR-21检测范围为0.1至200 pM的线性关系。结果表明,NMOF可作为有前景的核酸提取和荧光传感平台。

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