Electrochemical micro-aptasensors for exosome detection based on hybridization chain reaction amplification.
Electrochemical micro-aptasensors for exosome detection based on hybridization chain reaction amplification.
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DOI：
10.1038/s41378-021-00293-8
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发表时间：
2021
影响因子：
7.9
通讯作者：
Huang TJ
中科院分区：
文献类型：
--
作者：
Zhang W;Tian Z;Yang S;Rich J;Zhao S;Klingeborn M;Huang PH;Li Z;Stout A;Murphy Q;Patz E;Zhang S;Liu G;Huang TJ
外来体是细胞衍生的纳米囊泡，由于它们携带大量的分子货物，例如核酸和蛋白质，它们最近作为液体活检中的潜在生物标志物而受到欢迎。然而，大多数现有的基于外泌体的分析传感方法难以同时实现高灵敏度和高选择性。在这项工作中，我们提出了一种电化学微适体传感器的高度灵敏的检测外泌体集成的微图案化的电化学适体传感器和杂交链反应（HCR）信号放大方法。具体地，外泌体在CD 63适体功能化的电极上富集，然后被具有使用EpCAM适体作为桥连接的抗生物素蛋白-辣根过氧化物酶（HRP）的HCR产物识别。随后，通过HRP酶和3，3 '，5，5'-四甲基联苯胺（TMB）/H2 O2之间的酶反应产生的电流信号与HCR产物上结合的HRP的量直接相关，从而与靶外来体的数量直接相关。通过引入抗EpCAM适体，微适体传感器可以以高特异性检测癌性外泌体。由于微图案化电极和HCR双扩增策略，微适体传感器对于2.5×103至1×107个外泌体/mL的宽范围外泌体浓度实现线性检测响应，检测限为5×102个外泌体/mL。此外，我们的方法成功地检测了早期和晚期肺癌患者血清样品中的肺癌外泌体，展示了早期癌症诊断的巨大潜力。
Exosomes are cell-derived nanovesicles that have recently gained popularity as potential biomarkers in liquid biopsies due to the large amounts of molecular cargo they carry, such as nucleic acids and proteins. However, most existing exosome-based analytical sensing methods struggle to achieve high sensitivity and high selectivity simultaneously. In this work, we present an electrochemical micro-aptasensor for the highly sensitive detection of exosomes by integrating a micropatterned electrochemical aptasensor and a hybridization chain reaction (HCR) signal amplification method. Specifically, exosomes are enriched on CD63 aptamer-functionalized electrodes and then recognized by HCR products with avidin-horseradish peroxidase (HRP) attached using EpCAM aptamers as bridges. Subsequently, the current signal that is generated through the enzyme reaction between the HRP enzyme and 3,3’,5,5’-tetramethylbenzidine (TMB)/H2O2 directly correlates to the amount of bound HRP on the HCR products and thus to the number of target exosomes. By introducing anti-EpCAM aptamers, micro-aptasensors can detect cancerous exosomes with high specificity. Due to the micropatterned electrodes and HCR dual-amplification strategy, the micro-aptasensors achieve a linear detection response for a wide range of exosome concentrations from 2.5×103 to 1×107 exosomes/mL, with a detection limit of 5×102 exosomes/mL. Moreover, our method successfully detects lung cancer exosomes in serum samples of early-stage and late-stage lung cancer patients, showcasing the great potential for early cancer diagnosis.
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DOI：
10.1073/pnas.0407024101
发表时间：
2004-10-26
影响因子：
11.1
作者：
Dirks, RM;Pierce, NA
通讯作者：
Pierce, NA
影响因子：
64.5
作者：
Luga, Valbona;Zhang, Liang;Wrana, Jeffrey L.
通讯作者：
Wrana, Jeffrey L.
影响因子：
7.4
作者：
Luo X;An M;Cuneo KC;Lubman DM;Li L
通讯作者：
Li L
影响因子：
21.3
作者：
Skog, Johan;Wuerdinger, Tom;van Rijn, Sjoerd;Meijer, Dimphna H.;Gainche, Laura;Sena-Esteves, Miguel;Curry, William T., Jr.;Carter, Bob S.;Krichevsky, Anna M.;Breakefield, Xandra O.
通讯作者：
Breakefield, Xandra O.
影响因子：
7.4
作者：
Doldan, Ximena;Fagundez, Pablo;Tosar, Juan Pablo
通讯作者：
Tosar, Juan Pablo