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基于 RNA 切割 DNA zyme 的新型高灵敏组装-解离荧光传感策略用于 MDA-MB-231 乳腺癌生物标志物测定。

A novel highly sensitive compilation-detachment fluorescence sensing strategy based on RNA-cleavage DNAzyme for MDA-MB-231 breast cancer biomarker determination.

机构信息

Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj, Iran.

Nanotechnology Research Center, University of Kurdistan, P.O. Box 416, Sanandaj, Iran.

出版信息

J Mater Chem B. 2023 Feb 15;11(7):1568-1579. doi: 10.1039/d2tb02467e.

DOI:10.1039/d2tb02467e
PMID:36722940
Abstract

Herein, we designed a novel and highly sensitive fluorescence multicomponent detachable platform for MDA-MB-231 breast cancer cell detection as a model. The RNA cleavage DNAzyme was used as a central operator of the multicomponent probe through which compilation and induced detachment of probe was done. During the compilation step, the dsDNA-Sybr green 1 complexes on gold nanoparticles (GNP@dsDNA@SG1) were assembled. The intercalated Sybr green in the DNA structure has been used as an amplified signal generator on one site of DNAzyme and magnetic nanoparticles (MNP) act as a biological carrier and probe collector on the opposite side. The enzyme activator co-factor (MDA-MB-231 cell cytoplasmic protein) provokes the activation of the catalytic core of enzyme sequence in the DNAzyme molecule, followed by cleavage reaction in the substrate sequence and releasing GNP@ dsDNA@SG1 into the solution. The results indicate that the Sybr green emission fluorescence (520 nm) increases with the increment of MDA-MB-231 protein concentration in the linear dynamic range of 8.10 × 10 to 1.95 ng ml (0.77 × 10-0.019 cell ml) with a detection limit (LOD) of 1/72 × 10 pg ml under optimal conditions. The proposed immunosensor has great potential in developing ultrasensitive and rapid diagnostic platforms.

摘要

在这里,我们设计了一种新型的、高灵敏度的荧光多组分可分离平台,用于 MDA-MB-231 乳腺癌细胞检测作为模型。RNA 切割 DNA 酶被用作多组分探针的中心操作子,通过它完成探针的组装和诱导分离。在组装步骤中,金纳米粒子 (GNP@dsDNA@SG1) 上组装了双链 DNA-Sybr green 1 复合物。DNA 结构中的嵌入 Sybr green 被用作 DNA 酶和磁性纳米粒子 (MNP) 一侧的一个位点上的扩增信号发生器,MNP 作为生物载体和探针收集器在另一侧。酶激活辅因子 (MDA-MB-231 细胞细胞质蛋白) 引发 DNA 酶分子中酶序列的催化核心的激活,随后在底物序列中发生切割反应,并将 GNP@dsDNA@SG1 释放到溶液中。结果表明,随着 MDA-MB-231 蛋白浓度在 8.10×10 到 1.95ng ml (0.77×10-0.019 细胞 ml) 的线性动态范围内的增加,Sybr green 发射荧光 (520nm) 增加,在最佳条件下,检测限 (LOD) 为 1/72×10 pg ml。该免疫传感器在开发超灵敏和快速诊断平台方面具有巨大潜力。

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