Chen Yulin, Gao Dan, Zhu Qingyun, Chu Bizhu, Peng Jie, Wang Jian, Liu Liping, Jiang Yuyang
State Key Laboratory of Chemical Oncogenomics, Guangdong Provincial Key Laboratory of Chemical Biology, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, Guangdong, 518055, China.
The First Affiliated Hospital, Cancer Research Institute, Hengyang Medical School, University of South China, Hengyang 421001, China.
Analyst. 2023 May 16;148(10):2387-2394. doi: 10.1039/d3an00335c.
Exosomes are considered as promising biomarkers for early cancer diagnosis and prognosis. However, the majority of the research studies focused on a single type of exosomal biomarkers, which cannot comprehensively reflect the state of cancer for accurate diagnosis. To address this problem, we presented a ship-shaped microfluidic device containing a microcolumn array for simultaneous detection of exosomal surface proteins and miRNAs. Exosomes were first captured in the microchannels modified with CD63 protein aptamer. Exosomal surface proteins and miRNAs were simultaneously detected in four parallel channels to avoid the interference of fluorescent signals using specific aptamers labeled by Cy5 and catalytic hairpin assembly (CHA) based signal amplification strategy. The limit of detection for multiplexed markers in exosomes was 83 exosomes per μL, which is comparable to previously reported methods. Through quantitative analysis of two disease-specific surface proteins and miRNAs derived from different cancer cells and clinical serum samples, different cancer subtypes as well as cancer patients and healthy people could be significantly distinguished. These results suggest that this simple, highly sensitive, and more accurate analytical strategy by simultaneous profiling of different types of exosomal biomarkers has potential applications in cancer diagnosis and stage monitoring.
外泌体被认为是早期癌症诊断和预后的有前景的生物标志物。然而，大多数研究集中在单一类型的外泌体生物标志物上，这无法全面反映癌症状态以进行准确诊断。为了解决这个问题，我们提出了一种船形微流控装置，其包含用于同时检测外泌体表面蛋白和微小RNA（miRNA）的微柱阵列。外泌体首先被捕获在用CD63蛋白适配体修饰的微通道中。使用由Cy5标记的特异性适配体和基于催化发夹组装（CHA）的信号放大策略，在外泌体表面蛋白和miRNA在四个平行通道中同时被检测，以避免荧光信号的干扰。外泌体中多重标志物的检测限为每微升83个外泌体，这与先前报道的方法相当。通过对源自不同癌细胞和临床血清样本的两种疾病特异性表面蛋白和miRNA进行定量分析，可以显著区分不同的癌症亚型以及癌症患者和健康人。这些结果表明，通过同时分析不同类型的外泌体生物标志物，这种简单、高度灵敏且更准确的分析策略在癌症诊断和分期监测中具有潜在应用。
