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

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 (andgt;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.PROJECT NARRATIVE Extracellular vesicles (EVs) are small lipid bilayer particles secreted by most cell types. Profiling of EV-derived biomarkers provides a promising approach for diagnosis of various diseases. However, one of the primary hindrances to the study and application 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. To overcome the critical biological and technical hurdles, we propose to develop, optimize, and validate a Label-free Extracellular-vesicle Automated Purification System (LEAPS) with excellent EV-isolation capability over the other technologies.