School of Chemistry and Molecular Engineering, East China Normal University, 500 Dongchuan Road, Shanghai 200241, P. R. China.
Anal Chem. 2020 Apr 7;92(7):5404-5410. doi: 10.1021/acs.analchem.0c00106. Epub 2020 Mar 18.
Breast cancer is a heterogeneous disease, and it lacks special tumor markers. Exosomes, new noninvasive biomarkers, with the proteins on the exosome surface show potential for the diagnosis and prognosis of a tumor. However, assessing the variations of exosomal proteins still faces significant challenges. Herein, a magneto-mediated electrochemical sensor based on host-guest recognition has been developed for simultaneous analysis of breast cancer exosomal proteins. Magnetic beads (MB) modified with CD63 aptamer was first employed to capture exosomes. Silica nanoparticles (SiO NPs) was modified with MUC1, HER2, EpCAM, and CEA aptamers for specific exosomal proteins identification, respectively, and functionalized with -(2-((2-aminoethyl)disulfanyl)ethyl) ferrocene carboxamide (FcNHSSNH) as the signal molecule. The sandwich structure (MB-exosomes-SiO NPs probe) was separated by a magnet, and -(2-mercaptoethyl) ferrocene carboxamide (FcNHSH) was released to the supernatant by the addition of reductants (dithiothreitol, DTT) that break the disulfide bond of FcNHSSNH. FcNHSH and the graphene oxide-cucurbit 7 modified screen-printed carbon electrode (SPCE) was employed to monitor the oxidation current signals. In this way, four tumor markers on different breast cancer cells (MCF-7, SK-BR-3, MDA-MB-231, and BT474) derived exosomes were sensitively detected. Furthermore, the present assay enabled accurate analysis of exosomes from breast cancer patients, suggesting the potential of exosome analysis in clinic diagnosis.
乳腺癌是一种异质性疾病,缺乏特殊的肿瘤标志物。外泌体作为新型的非侵入性生物标志物,其表面的蛋白质显示出在肿瘤诊断和预后方面的潜力。然而,评估外泌体蛋白的变化仍然面临着重大的挑战。在此,我们开发了一种基于主体-客体识别的磁介导电化学传感器,用于同时分析乳腺癌外泌体蛋白。首先,用 CD63 适体修饰的磁珠(MB)来捕获外泌体。然后,用 MUC1、HER2、EpCAM 和 CEA 适体分别修饰硅纳米粒子(SiO NPs),用于特异性外泌体蛋白的识别,并将 -(2-((2-氨乙基)二硫代)乙基)二茂铁羧酰胺(FcNHSSNH)功能化作为信号分子。通过磁铁分离三明治结构(MB-外泌体-SiO NPs 探针),并用还原剂(二硫苏糖醇,DTT)加入来释放到上清液中的 -(2-巯基乙基)二茂铁羧酰胺(FcNHSH),还原二硫键 FcNHSSNH。FcNHSH 和氧化石墨烯-瓜环[7](GO-CB[7])修饰的丝网印刷碳电极(SPCE)被用来监测氧化电流信号。通过这种方式,我们可以灵敏地检测来自不同乳腺癌细胞(MCF-7、SK-BR-3、MDA-MB-231 和 BT474)的外泌体中的四种肿瘤标志物。此外,本研究能够准确分析乳腺癌患者的外泌体,提示外泌体分析在临床诊断中的应用潜力。