Nanopore Array for Multiparameter Analysis of Single Extracellular Vesicles

项目摘要 今天，学术界和工业界的研究人员和临床医生缺乏必要的工具和技术来 准确地表征、区分和分类细胞外小泡(EV)；目前可用的技术包括 根本不是用来描述尺寸小于200纳米的实体的。虽然电动汽车的重要性 在引导用于蜂窝间通信的货物递送时，无法描述真实的分布 对其生物物理性质的了解是了解其作用机制、鉴定 疾病特异性生物标记物，以及利用以肠道病毒为基础的纳米药物的治疗潜力。在此期间 电子生物科学(EBS)第一阶段STTR计划，一家拥有成功记录的公司 将基于纳米孔的分析技术和相关的高性能、低噪声 将与犹他大学杰出教授亨利·S·怀特合作，后者是 纳米孔、质量传输和电分析表征方面的顶尖专家，开发和演示 一种全新的技术，能够进行最先进的、全面的电动汽车表征和样本分析 在单泡水平上。此外，快速评估单个电动汽车的能力将使分类能力成为可能 并分离特定的EV亚群，以供进一步下游表征和/或利用。在此期间 项目，将建立一个原型系统，并对综合表征和分类/分离的可行性 将对特定的EV人群进行演示和验证。由此产生的系统将具有易用性， 吞吐量，以及高通量、低分辨率流式细胞仪的价位，但状态 最先进的显微技术，例如扫描或透射式电子显微镜(扫描电子显微镜或电子显微镜， 分别)或原子力显微镜(AFM)。提议的技术的优势将是 开发一个系统，使其能够识别和分类/隔离精确定义的 使用高维(多参数)数据的具有生物和医学意义的纳米级(200 Nm)实体， 它的应用将远远超出电动汽车，包括但不限于表征其他纳米结构 在药物输送和基因治疗中很重要的纳米药物，如疫苗用脂质纳米粒，外切体- 基于治疗的药物、慢病毒、病毒样颗粒和基于聚合物的纳米胶囊。因此，成功的 这一计划的完成将导致一个广泛适用的工具，能够将精度施加在这些最小的 并实现了推进电动汽车和电动汽车领域亟需的表征/分类 纳米医学。

Project Summary Today, researchers and clinicians, in academia and industry, lack the tools and technologies needed to accurately characterize, differentiate, and sort extracellular vesicles (EVs); currently available technologies were simply not developed to characterize entities smaller than 200 nanometers in size. While the importance of EVs in directing cargo delivery for intercellular communication is well known, the inability to profile the true distribution of their biophysical properties is the main hindrance to understanding the mechanisms of action, identifying disease-specific biomarkers, and harnessing the therapeutic potential of EV-based nanomedicine. During this Phase I STTR program, Electronic BioSciences (EBS), a company with a successful track record of commercializing nanopore-based analytical technology and the associated high-performance, low-noise electronics, will collaborate with Distinguished Professor Henry S. White at the University of Utah, who is a foremost expert in nanopores, mass transport and electroanalytical characterization, to develop and demonstrate an entirely new technology capable of state-of-the-art, comprehensive EV characterization and sample profiling at the single-vesicle level. Furthermore, the ability to quickly assess individual EVs will enable the ability to sort and isolate specific EV subpopulations for further downstream characterization and/or utilization. During this project, a prototype system will be built and the feasibility of comprehensively characterizing and sorting/isolating specific EV populations will be demonstrated and validated. The resulting system will have the ease-of-use, throughput, and price point of a high-throughput, “low-resolution” flow-cytometer, but the resolution of state-of- the-art microscopy techniques, such as scanning or transmission electron microscopy (SEM or TEM, respectively) or atomic force microscopy (AFM). The advantage of the proposed technology will be the development of a system enabling the ability to identify and sort/isolate precisely defined populations of biological- and medical-significance nanoscale (<200 nm) entities using high-dimensional (multiparameter) data, which will have applications far beyond EVs, including but not limited to characterizing other nanostructures important in drug delivery and gene therapy nanomedicines such as lipid nanoparticles for vaccines, exosome- based therapeutics, lentivirus, virus-like particles, and polymer-based nanocapsules. Thus, the successful completion of this program will result in a broadly applicable tool able to bring precision to bear on these smallest of couriers and enable the characterization/classification urgently needed to advance the fields of EVs and nanomedicine.