Label-Free Small Extracellular Vesicles Capturing Strategy for Lung Cancer Diagnosis and Typing Based on a Natural Polyphenol-Metal Three-Dimensional Network
Label-Free Small Extracellular Vesicles Capturing Strategy for Lung Cancer Diagnosis and Typing Based on a Natural Polyphenol-Metal Three-Dimensional Network
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DOI：
10.1021/acs.analchem.2c03283
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发表时间：
2022-11-10
影响因子：
7.4
通讯作者：
Qiao, Xiaoqiang
中科院分区：
文献类型：
--
作者：
Wang, Haiyan;Liu, Delu;Qiao, Xiaoqiang
细胞外小泡（Small extracellular vesicles，sEVs）作为循环生物标志物和疾病诊断的指示剂已越来越受到重视。然而，sEV的临床应用受到缺乏稳健且易于放大的分离技术的严重限制。本文中，探索了用于无标记sEV分离的多酚-金属三维（3D）网络的可行性。作为概念验证，以单宁酸（TA）为多酚配体，Fe（III）为配位金属，通过配位驱动的层层自组装方法，设计并制备了TA-Fe（III）三维网络包覆的介孔二氧化硅微球（SiO2@ BSA@ Fe-TA 6）。通过傅里叶变换红外光谱、扫描电子显微镜、X射线光电子能谱和热重分析验证了SiO2@BSA@ Fe-TA 6的成功制备。以低成本TA（低至0.18美元/g）为探针，SiO2@BSA@ Fe-TA 6实现了对不同细胞和血浆样品中sEV的通用捕获。捕获效率达到85.4 +/-1.5%，这与基于抗体的捕获技术相当，并且显著高于超离心（UC）方法。通过SiO2@BSA@ Fe-TA 6从H1299细胞培养上清液中分离的sEV的纯度测量为（1.07 +/- 0.14）× 1011个颗粒/μ g，这是通过UC方法的3.1倍。SiO2 @ BSA @ Fe-TA 6的另一个重要优势是简单的自组装方法，可以收获高达克的产量，允许同时处理500多个血浆样品。基于SiO2@BSA@ Fe-TA 6的策略进一步成功地用于区分非小细胞肺癌（NSCLC）和小细胞肺癌（SCLC），准确率为87.1%。所开发的SiO2@BSA@ Fe-TA 6是一种用于肺癌诊断和分型中的基于sEV的液体活检的无标记、通用、低成本且易于放大的技术。
Small extracellular vesicles (sEVs) have been increasingly recognized as circulating biomarkers and prognostica-tors for disease diagnosis. However, the clinical applications of sEVs are seriously limited by the lack of a robust and easy scale-up isolation technique. Herein, the feasibility of a polyphenol-metal three-dimensional (3D) network for label-free sEV isolation was explored. As a proof-of-concept, with tannic acid (TA) as the polyphenolic ligand and Fe(III) as the coordinated metal, the TA-Fe(III) 3D network coating mesoporous silica beads (SiO2@ BSA@Fe-TA6) was designed and fabricated via a coordination-driven layer-by-layer self-assembly approach. The successful fabrication of SiO2@BSA@Fe-TA6 was validated by Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray photoelectron spectroscopy, and thermogravimetric analysis. With the low-cost TA (as low as US$ 0.18/g) as the probe, SiO2@BSA@Fe-TA6 achieved universal capture toward sEVs in different cells and plasma samples. The capture efficiency reached 85.4 +/- 1.5%, which is comparable to the antibody-based capture techniques and significantly higher than the ultracentrifugation (UC) method. The purity of sEVs isolated by SiO2@BSA@Fe-TA6 from the H1299 cell culture supernatant was measured as (1.07 +/- 0.14) x 1011 particles/mu g, which is 3.1 times higher than that via the UC method. Another important superiority of SiO2@BSA@ Fe-TA6 is the facile self-assembly approach, which can harvest a yield of up to grams, allowing simultaneous processing of more than 500 plasma samples. The SiO2@BSA@Fe-TA6-based strategy was further successfully employed to distinguish nonsmall cell lung cancer (NSCLC) and small cell lung cancer (SCLC) with an accuracy of 87.1%. The developed SiO2@BSA@Fe-TA6 is a label-free, universal, low cost, and easy scale-up technique for sEV-based liquid biopsy in lung cancer diagnosis and typing.