An integrative microfluidic device for isolation and ultrasensitive detection of lung cancer-specific exosomes from patient urine
An integrative microfluidic device for isolation and ultrasensitive detection of lung cancer-specific exosomes from patient urine
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用于从患者尿液中分离和超灵敏检测肺癌特异性外泌体的集成微流体装置
DOI：
10.1016/j.bios.2020.112290
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发表时间：
2020-09-01
影响因子：
12.6
通讯作者：
Liu, Fei
中科院分区：
文献类型：
--
作者：
Yang, Qinsi;Cheng, Liming;Liu, Fei
来源于各种生物流体的外泌体可以作为癌症早期检测的潜在生物标志物。然而，由于缺乏可靠的分离和检测方法，外泌体临床翻译仍然具有挑战性。在此，我们提出了一种新的集成微流体装置，专门设计用于从患者尿液中收集的肺癌特异性外泌体的分离和原位检测。这种新装置是用聚甲基丙烯酸甲酯（PMMA）和用捕获抗体修饰的纳米多孔金（Au）纳米簇膜制造的。然后加载第二抗体缀合的Au纳米棒探针以使用暗视野显微镜鉴定和定量肺癌特异性外泌体。由于共振瑞利散射，AuNC-外泌体-AuR复合物产生显著的散射波长偏移和改善的散射强度，这使得能够以低于1000个颗粒/mL的LOD超灵敏地检测外泌体。蛋白质组学分析表明，高纯度的外泌体已被分离的设备。然后，我们用500 μ L肺癌患者和对照组的尿液样本验证了这种方法，这对区分早期肺癌患者和健康个体表现出很大的希望。总之，所提出的方法是快速和超灵敏的，并且可以容易地适用于从其他恶性肿瘤中分离和检测癌症特异性外泌体。
Exosomes, derived from various biofluids, may serve as potential biomarkers for cancer early detection. Nevertheless, exosome clinical translation remains challenging due to the lack of reliable isolation and detection methods. Herein, we present a novel integrated microfluidic device specifically designed for isolation and in-situ detection of lung cancer-specific exosomes collected from patient's urine. The new device has been fabricated using polymethyl methacrylate (PMMA) and a nanoporous gold (Au) nanocluster membrane modified with the capture antibody. The second antibody-conjugated Au nanorod probe was then loaded to identify and quantify lung cancer-specific exosomes using a dark field microscope. AuNC-Exosome-AuR complex produces a significant scattering wavelength shift and an improved scattering intensity due to resonance Rayleigh scattering, which enables the ultrasensitive detection of exosomes with a LOD below 1000 particles/mL. The proteomic analysis revealed that the high-purity exosomes has been isolated by the device. We then validated this method with 500 mu L urine samples from lung cancer patients and controls, which showed great promise for differentiating early-stage lung cancer patients from healthy individuals. Taken together, the presented method is fast and ultrasensitive and can be easily adapted for the isolation and detection of cancer specific exosomes from other malignant tumors.