State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha, 410082, PR China.
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha, 410082, PR China.
Anal Chim Acta. 2023 May 8;1254:341130. doi: 10.1016/j.aca.2023.341130. Epub 2023 Mar 22.
Exosomes, as a non-invasive biomarker, perform an important role in breast cancer screening and prognosis monitoring. However, establishing a simple, sensitive, and reliable exosome analysis technique remains challenging. Herein, a one-step multiplex analysis electrochemical aptasensor based on a multi-probe recognition strategy was constructed to analyze breast cancer exosomes. HER2-positive breast cancer cell (SK-BR-3) exosomes were selected as the model targets and three aptamers including CD63, HER2 and EpCAM aptamers were used as the capture units. Methylene blue (MB) functionalized HER2 aptamer and ferrocene (Fc) functionalized EpCAM aptamer, which were modified on gold nanoparticles (Au NPs), i.e. MB-HER2-Au NPs and Fc-EpCAM-Au NPs, were used as signal units. When the mixture of target exosomes, MB-HER2-Au NPs and Fc-EpCAM-Au NPs were added on the CD63 aptamer modified gold electrode, two Au NPs modified by MB and Fc could be specifically captured on the electrode by the recognition of three aptamers with target exosomes. Then one-step multiplex analysis of exosomes was achieved by detecting two independent electrochemical signals. This strategy can not only distinguish breast cancer exosomes from other exosomes (including normal exosomes and other tumor exosomes) but also HER2-positive breast cancer exosomes and HER2-negative breast cancer exosomes. Besides, it had high sensitivity and can detect SK-BR-3 exosomes with a concentration as low as 3.4 × 10 particles mL. Crucially, this method can be applicable to the examination of exosomes in complicated samples, which is anticipated to afford assistance for the screening and prognosis of breast cancer.
外泌体作为一种非侵入性生物标志物，在乳腺癌筛查和预后监测中发挥着重要作用。然而，建立一种简单、灵敏、可靠的外泌体分析技术仍然具有挑战性。在此，构建了一种基于多探针识别策略的一步式多重分析电化学适体传感器，用于分析乳腺癌外泌体。选择 HER2 阳性乳腺癌细胞（SK-BR-3）外泌体作为模型靶标，使用三种适体作为捕获单元，包括 CD63、HER2 和 EpCAM 适体。将亚甲基蓝（MB）功能化的 HER2 适体和二茂铁（Fc）功能化的 EpCAM 适体修饰在金纳米粒子（Au NPs）上，即 MB-HER2-Au NPs 和 Fc-EpCAM-Au NPs，用作信号单元。当将靶向外泌体、MB-HER2-Au NPs 和 Fc-EpCAM-Au NPs 的混合物加入到 CD63 适体修饰的金电极上时，两个修饰有 MB 和 Fc 的 Au NPs 可以通过与靶向外泌体的三个适体的识别特异性地被捕获在电极上。然后，通过检测两个独立的电化学信号，实现了对外泌体的一步式多重分析。该策略不仅可以区分乳腺癌外泌体与其他外泌体（包括正常外泌体和其他肿瘤外泌体），还可以区分 HER2 阳性乳腺癌外泌体和 HER2 阴性乳腺癌外泌体。此外，它具有高灵敏度，能够检测浓度低至 3.4×10 个粒子/mL 的 SK-BR-3 外泌体。至关重要的是，该方法可适用于复杂样本中外泌体的检查，有望为乳腺癌的筛查和预后提供帮助。
