Aqueous two phase system microfluidic isolation and concentration of extracellular vesicles

这项NSERC创新理念（I2 I）资助正在资助微流体平台技术的开发，该技术可以直接从体液（如全血和尿液）中分离和纯化细胞外囊泡（EV）。这种微流体方法导致EV浓度比使用当前金标准超离心技术可实现的浓度高几倍。我们的方法结合了微流体的精确性与水两相系统（ATPS）流体的选择性分配能力，以分离和纯化EV，重要的是，克服了超离心的关键限制-即EV浓度太低，无法应用，杂质共分离和EV损伤。我们技术的关键创新是微流体和内部优化的ATPS的结合，该ATPS利用热力学能量平衡将EV轻轻地穿梭到一个流体相，同时将所有其他体液成分移动到不同的流体相，并增加了第二种方法来选择特定的EV亚群。该项目的具体目标是：1）针对特定的EV亚群优化ATPS，2）将ATPS集成到微流体格式中，以实现最大的EV分区。一家位于温哥华的生物技术公司已经被该项目所吸引，因为该项目在生物医学领域的潜在影响。该项目的完成将使我们能够将该技术授权给潜在的商业合作伙伴，并实现一种新的EV分离和纯化工具，该工具在EV研究中具有重要用途，并使液体活检广泛用于癌症诊断和治疗监测应用。

This NSERC Idea to Innovation (I2I) grant is funding the development of a microfluidic platform technology that directly isolates and purifies extracellular vesicles (EVs) from bodily fluids, like whole blood and urine. This microfluidic approach results in EV concentrations that are several times higher than the concentrations achievable using the current gold-standard ultracentrifugation technique. Our approach combines the precision of microfluidics with the selective partitioning capability of aqueous two-phase system (ATPS) fluids, to isolate and purify EVs, and importantly, overcome the critical limitations of ultracentrifugation--namely, EV concentrations that are too low for applications, co-isolation of impurities, and EV damage. The key innovations of our technology are the marriage of microfluidics and an in-house optimized ATPS that exploits thermodynamic energy balances to gently shuttle EVs to one fluid phase, while moving all other bodily fluid components to a different fluid phase, and the addition of a secondary approach to select for specific EV subpopulations. The specific objectives in this project are to 1) optimize an ATPS for specific subpopulations of EVs, and 2) integrate the ATPS into a microfluidic format for maximum EV partitioning.A Vancouver-based biotechnology company is already attracted to this project because of the project's potential impact in the area of biomedicine. The completion of this project will lead us to the position of being able to license the technology to potential commercial partners, and realize a new EV isolation and purification tool that has important utility in EV research, and makes liquid biopsies widely usable in cancer diagnosis and treatment monitoring applications.