Rapid and unbiased enrichment of extracellular vesicles via a meticulously engineered peptide

IF 18 1区 医学 Q1 ENGINEERING, BIOMEDICAL
Le Wang , Zhou Gong , Ming Wang , Yi-Zhong Liang , Jing Zhao , Qi Xie , Xiao-Wei Wu , Qin-Ying Li , Cong Zhang , Li-Yun Ma , Si-Yang Zheng , Ming Jiang , Xu Yu , Li Xu
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Abstract

Extracellular vesicles (EVs) have garnered significant attention in biomedical applications. However, the rapid, efficient, and unbiased separation of EVs from complex biological fluids remains a challenge due to their heterogeneity and low abundance in biofluids. Herein, we report a novel approach to reconfigure and modify an artificial insertion peptide for the unbiased and rapid isolation of EVs in 20 min with ∼80% recovery in neutral conditions. Moreover, the approach demonstrates exceptional anti-interference capability and achieves a high purity of EVs comparable to standard ultracentrifugation and other methods. Importantly, the isolated EVs could be directly applied for downstream protein and nucleic acid analyses, including proteomics analysis, exome sequencing analysis, as well as the detection of both epidermal growth factor receptor (EGFR) and V-Ki-ras2 Kirsten Rat Sarcoma Viral Oncogene Homologue (KRAS) gene mutation in clinical plasma samples. Our approach offers great possibilities for utilizing EVs in liquid biopsy, as well as in various other biomedical applications.

Abstract Image

通过精心设计的多肽快速、无偏见地富集细胞外囊泡
细胞外囊泡(EVs)在生物医学应用中备受关注。然而,由于EVs在生物流体中的异质性和低丰度,从复杂的生物流体中快速、高效、无偏见地分离EVs仍然是一项挑战。在此,我们报告了一种重组和修饰人工插入肽的新方法,该方法可在 20 分钟内无偏见地快速分离 EVs,中性条件下的回收率可达 80%。此外,该方法还显示出卓越的抗干扰能力,实现了与标准超速离心法和其他方法相当的高纯度 EVs。重要的是,分离出的EVs可直接用于下游蛋白质和核酸分析,包括蛋白质组学分析、外显子组测序分析以及临床血浆样本中表皮生长因子受体(EGFR)和V-Ki-ras2 Kirsten鼠肉瘤病毒同源物(KRAS)基因突变的检测。我们的方法为在液体活检以及其他各种生物医学应用中利用 EVs 提供了极大的可能性。
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来源期刊
Bioactive Materials
Bioactive Materials Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
28.00
自引率
6.30%
发文量
436
审稿时长
20 days
期刊介绍: Bioactive Materials is a peer-reviewed research publication that focuses on advancements in bioactive materials. The journal accepts research papers, reviews, and rapid communications in the field of next-generation biomaterials that interact with cells, tissues, and organs in various living organisms. The primary goal of Bioactive Materials is to promote the science and engineering of biomaterials that exhibit adaptiveness to the biological environment. These materials are specifically designed to stimulate or direct appropriate cell and tissue responses or regulate interactions with microorganisms. The journal covers a wide range of bioactive materials, including those that are engineered or designed in terms of their physical form (e.g. particulate, fiber), topology (e.g. porosity, surface roughness), or dimensions (ranging from macro to nano-scales). Contributions are sought from the following categories of bioactive materials: Bioactive metals and alloys Bioactive inorganics: ceramics, glasses, and carbon-based materials Bioactive polymers and gels Bioactive materials derived from natural sources Bioactive composites These materials find applications in human and veterinary medicine, such as implants, tissue engineering scaffolds, cell/drug/gene carriers, as well as imaging and sensing devices.
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