A Label-free Extracellular-vesicle Automated Purification System (LEAP System)

项目摘要/摘要 细胞外小泡(EV)是一种由大多数细胞类型分泌的小的脂质双层颗粒，它包裹着 生物分子，包括来自亲本细胞的脂类、蛋白质和核酸。肠道病毒衍生生物标志物的特征分析 为各种疾病的早期诊断和预后提供了一种很有前途的方法。目前，其中一家 阻碍电动汽车实施和使用的主要障碍是缺乏一种方便和高效的可靠方法。 快速分离出产量高、纯度高的电动汽车。最近的一些出版物提到了发展中的 高效、可靠的分离方法是该领域进一步发展的迫切需要。有一种强烈的探索欲望 在产量、纯度、产量、速度、 健壮性和成本。为了克服这些挑战，WellSIM Biomedical Technologies计划开发， 优化和验证电动汽车的无标签胞外囊泡自动净化系统(LEAP系统) 与体液(如血浆和尿液)隔离，性能优异。基于我们目前的原型 已经显示出具有竞争力的电动汽车产量、纯度和分离效率，我们将进一步改进加工 通过重新设计和优化系统来提高速度、样本量负载和吞吐量。我们的初步研究 支持，通过提议的改进，我们有望同时从多达6个血浆中分离出电动汽车 样品(&gt；2毫升)在1小时内纯度和产量均优于其他EV分离方法。系统升级后 和优化，我们将全面验证我们的下一代平台的细胞培养液，尿液， 和等离子体在产量、纯度、速度和加工量方面。我们的新平台将直接解决 目前的瓶颈，从而为基于电动汽车的生命科学和精确化提供了先进的电动汽车净化工具 医药。

PROJECT SUMMARY / ABSTRACT Extracellular vesicles (EVs) are small lipid bilayer particles secreted by most cell types, which encapsulates biomolecules including lipids, proteins, and nucleic acids from parent cells. Profiling of EV-derived biomarkers provides a promising approach for early diagnoses and prognosis of various diseases. Currently, one of the primary hindrances to the implementation and usage of EVs is lack of a robust method for convenient and high- speed isolation of EVs with high yield and purity. A number of recent publications mentioned that developing efficient and reliable isolation methods is urgent to further advance in this field. There is a strong desire to explore innovated technologies for better performance of EV isolation in terms of throughput, purity, yield, speed, robustness, and cost. To overcome these challenges, WellSIM Biomedical Technologies proposes to develop, optimize, and validate a Label-free Extracellular-vesicle Automated Purification System (LEAP System) for EV isolation from bodily fluids (e.g., plasma and urine) with excellent performance. Based on our current prototype which has shown competitive EV yield, purity, and isolation efficiency, we will further improve the processing speed, sample volume load, and throughput by redesigning and optimizing the System. Our preliminary studies support that, with the proposed improvement, we can expect to simultaneously isolate EVs from up to 6 plasma samples (>2 mL) within 1 hr with purity and yield outperform other EV isolation methods. After system upgrade and optimization, we will comprehensively validate our next-generation platform with cell culture medium, urine, and plasma in terms of yield, purity, speed, and processing volume. Our new platform will directly address the current bottlenecks, and thus provide an advanced EV purification tool for EV-based life science and precision medicine.