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基于微流控技术通过肽-纳米线混合界面捕获和释放癌症衍生外泌体

Microfluidic-based capture and release of cancer-derived exosomes via peptide-nanowire hybrid interface.

作者信息

Suwatthanarak Thanawat, Thiodorus Ivan Adiyasa, Tanaka Masayoshi, Shimada Taisuke, Takeshita Daiki, Yasui Takao, Baba Yoshinobu, Okochi Mina

机构信息

Department of Chemical Science and Engineering, Tokyo Institute of Technology, 2-12-1 O-okayama, Meguro-ku, Tokyo 152-8552, Japan.

出版信息

Lab Chip. 2021 Feb 7;21(3):597-607. doi: 10.1039/d0lc00899k. Epub 2020 Dec 24.

Abstract

Cancer-derived circulating exosomes or nanoscale extracellular vesicles are emerging biomarkers for disease detection and treatment because of their cell-specific constituents and unique intercellular pathways. For efficient exosome isolation from bio-fluids, the design of high-affinity nanointerfaces is of great importance in the development of miniaturized systems for the collection of exosomes. Herein, we report peptide-functionalized nanowires as a biorecognition interface for the capture and release of cancer-derived exosomes within a microfluidic channel. Based on the amino-acid sequence of EWI-2 protein, a partial peptide that bound to the CD9 exosome marker and thus targeted cancer exosomes was screened. Linkage of the exosome-targeting peptide with a ZnO-binding sequence allowed one-step and reagent-free peptide modification of the ZnO nanowire array. As a result of peptide functionalization, the exosome-capturing ability of ZnO nanowires was significantly improved. Furthermore, the captured exosomes could be subsequently released from the nanowires under a neutral salt condition for downstream applications. This engineered surface that enhances the nanowires' efficiency in selective and controllable collection of cancer-derived exosomes provides an alternative foundation for developing microfluidic platforms for exosome-based diagnostics and therapeutics.

摘要

癌症衍生的循环外泌体或纳米级细胞外囊泡因其细胞特异性成分和独特的细胞间途径,正成为疾病检测和治疗的生物标志物。为了从生物流体中高效分离外泌体,高亲和力纳米界面的设计对于开发用于外泌体收集的小型化系统至关重要。在此,我们报道了肽功能化纳米线作为一种生物识别界面,用于在微流控通道内捕获和释放癌症衍生的外泌体。基于EWI-2蛋白的氨基酸序列,筛选出了一种与CD9外泌体标志物结合从而靶向癌症外泌体的部分肽段。将外泌体靶向肽与ZnO结合序列相连,实现了ZnO纳米线阵列的一步法且无需试剂的肽修饰。肽功能化的结果是,ZnO纳米线捕获外泌体的能力显著提高。此外,在中性盐条件下,捕获的外泌体随后可从纳米线上释放出来用于下游应用。这种提高纳米线在选择性和可控收集癌症衍生外泌体方面效率的工程化表面,为开发基于外泌体的诊断和治疗微流控平台提供了另一种基础。

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