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

无标记细胞外囊泡自动纯化系统（LEAP系统）

• 批准号: 10324624
• 负责人: Yuchao Chen
• 金额: $ 19.75万
• 依托单位: WELLSIM BIOMEDICAL TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324624
• 关键词: Acoustics; Address; Adoption; Affinity; Biological; Biological Markers; Biological Sciences; Biomedical Technology; Blood Plasma Volume; Cell Culture Techniques; Cells; Chromatography; Clinical; Collaborations; Complex; Consumption; Data; Density Gradient Centrifugation; Development; Devices; Diagnosis; Diagnostic; Disease; Early Diagnosis; Encapsulated; Ensure; Evaluation; Goals; Knowledge; Label; Lipid Bilayers; Lipids; Liquid substance; Medicine; Membrane; Methods; Microfluidics; Nucleic Acids; Outcome; Parents; Performance; Phase; Plasma; Precipitation; Process; Prognosis; Proteins; Publications; Reagent; Recovery; Research; Research Personnel; Saliva; Sampling; Scientist; Small Business Innovation Research Grant; Source; Speed; System; Technology; Testing; Time; Transducers; Translations; Ultracentrifugation; Ultrafiltration; Urine; Work; base; cell type; clinical application; clinical translation; cost; cost effective; design; expectation; extracellular vesicles; flexibility; improved; innovation; innovative technologies; magnetic field; nanoparticle; new technology; next generation; novel; novel strategies; particle; precision medicine; pressure; processing speed; prototype; scale up; success; technological innovation; tool

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.

项目总结/摘要 细胞外囊泡是由大多数细胞分泌的小的脂质双层颗粒， 生物分子，包括来自亲本细胞的脂质、蛋白质和核酸。EV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有前途的方法。目前， 电动汽车的实施和使用的主要障碍是缺乏一种可靠的方法， 以高产率和高纯度快速分离EV。最近的一些出版物提到， 有效可靠的分离方法是该领域亟待进一步发展的。有强烈的探索欲望 创新技术，在产量、纯度、产量、速度 鲁棒性和成本。为了克服这些挑战，WellSIM生物医学技术公司建议开发， 优化和验证EV的无标记细胞外囊泡自动纯化系统（LEAP系统） 与体液隔离（例如，血浆和尿液），性能优异。基于我们现有的原型 已经显示出具有竞争力的EV产量、纯度和分离效率，我们将进一步改进处理 通过重新设计和优化系统，提高了系统的速度、样品体积负载和吞吐量。我们的初步研究 支持，与拟议的改进，我们可以期待同时隔离电动汽车从多达6个血浆 样品（>2 mL）在1小时内的纯度和产率优于其他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与细胞培养基，尿液， 和等离子体的产量、纯度、速度和处理量。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密制造提供先进的电动汽车净化工具。 药