A colorimetric aptasensor based on a hemin/EpCAM aptamer DNAzyme for sensitive exosome detection

适体 血红素 脱氧核酶 G-四倍体 外体 化学 检出限 微泡 生物化学 分子生物学 色谱法 生物 DNA 基因 小RNA 血红素
作者
Jingjing Kuang,Zhibo Fu,Xuezhi Sun,Chuhui Lin,Shenglong Yang,Jiayao Xu,Min Zhang,Hongyang Zhang,Fanghong Ning,Ping Hu
出处
期刊:Analyst [Royal Society of Chemistry]
卷期号:147 (22): 5054-5061 被引量:27
标识
DOI:10.1039/d2an01410f
摘要

Exosomes are considered as potential biomarkers that can reflect information from their parent cell-associated cancer microenvironment. Recently, aptasensors have been widely used for cancer and tumor exosome detection. Aptamers related to exosome surface proteins are usually used to introduce a sequence; the aptamer is used for exosome recognition, and the introduced sequence is used to form G-quadruplexes and for signal amplification. In this paper, we found that the EpCAM aptamer is rich in guanine and unimolecular G-quadruplex with a two-layer G-tetrad under acidic conditions, and we investigated its topology, thermal stability and dissociation constant with hemin. Based on this, our proposed colorimetric aptamer sensor combines the unmodified EpCAM aptamer with hemin to construct a hemin/G-quadruplex DNAzyme and catalyze the TMB-H2O2 system to generate a strong colorimetric signal. Therefore, colorimetric signal changes were negatively correlated with the exosome concentration. The linear range of the 1 h assay was 106-108 particles per mL, and the detection limit was 3.94 × 105 particles per mL. In addition, this method can detect exosomes in complex fetal bovine serum samples with good specificity and high sensitivity toward exosomes from breast, liver, and lung cancers with abnormal EpCAM protein expression.
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