Development of a high-yield and high-purity isolation kit for extracting extracellular vesicles from complex biological samples

拟议的研究项目包括圭尔夫大学(安大略省圭尔夫大学)的生物医学创新(BMI)实验室(李博士)和Galenvs Science Inc.(魁北克蒙特利尔)之间的合作伙伴关系。合作研究伙伴关系的主要目标是开发一种EV分离试剂盒，以高产率和高纯度从复杂的生物液体(如血浆/血清、尿液)中分离和纯化细胞外小泡(EV)。EVS是从细胞分泌到体液中的纳米级囊泡，如血液和尿液。由于EV携带的生物分子与其来源细胞相似，因此分析生物液中EV的含量已成为疾病诊断和治疗监测的一种有前途的工具。EVS(特别是外切体)也有能力作为治疗复杂疾病的平台，但免疫原性和致瘤性较低。然而，由于目前技术的产量和纯度的限制，从生物体液中分离EV构成了这些应用的主要瓶颈。本项目旨在通过开发一种aptamer-functionalized-magnetic-nanoparticle-based提取试剂盒来克服这些挑战，该试剂盒具有易于遵循的操作规程和即用即用的试剂，可从各种类型的生物液中分离EV，产率高，纯度高，适用于EV下游的分子分析。该项目的成功完成对加拿大的生物技术产业产生了重大的经济影响。潜在的成果和影响包括：(1)加拿大制造的首批EV隔离试剂盒之一，包括已制定的协议和即用试剂；(2)使EV在健康和生命科学中得到更广泛的应用；(3)提高加拿大生物技术公司在国内和全球市场的竞争力；(4)培训HQP纳米生物技术和EV科学。

The proposed research project includes a partnership between the BioMed Innovation (BMI) laboratory (Dr. Li) at the University of Guelph (UofG, Guelph, Ontario) and Galenvs Sciences Inc. (Montreal, Quebec). The primary goal of the collaborative research partnership is to develop an EV isolation kit to isolate and purify extracellular vesicles (EVs) from complex biological fluids (e.g., blood plasma/serum, urine) with high yield and high purity. EVs are nanoscale vesicles secreted from cells into body fluids such as blood and urine. Since EVs carry biomolecules similar to their cells of origin, analysis of EV contents in biofluids has been a promising tool for disease diagnosis and treatment monitoring. EVs (specifically, exosomes) also have the capacity to serve as a therapeutic platform to treat complex diseases, with low immunogenicity and tumorigenicity. However, EV isolation from biofluids constitutes a major bottleneck in these applications largely due to the yield and purity limitations of the current technologies. This project aims to overcome these challenges by developing an aptamer-functionalized-magnetic-nanoparticle-based extraction kit with easy-to-follow protocols and ready-to-use reagents for isolating EVs from various types of biofluids, with high yield and purity, suitable for downstream EV molecular analysis. Successful completion of this project has a substantial economic impact on the biotech industry of Canada. The potential outcomes and impacts include: (1) one of the first EV isolation kits made in Canada with both established protocols and ready-to-use reagents; (2) enabling broader applications of EVs in health and life sciences; (3) enhancing the competitiveness of the Canadian biotechnology companies both at home and in the global market; (4) training of HQP in nano-biotechnology and EV science.