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Elevating & Automating Cell and Extracellular Vesicle Purification with Digital Magnetic Sorting

Burwell, A., Bshara-Corson, S., Ayala, D. Ramirez, S., Tiemann, T.,, Murray, C. “ Elevating & Automating Cell and Extracellular Vesicle Purification with Digital Magnetic Sorting” Society of Laboratory and Automation & Screening Microscale Symposium." September 14, 2023. San Diego, CA, USA.

Precision diagnostics is experiencing a significant transformation with the emergence of extracellular vesicles (EVs) as significant biomarkers for multiple disease indications. Among these, immune cell-derived EVs have shown potential as biomarkers for Graft vs Host Disease (GVHD) as they exhibit a strong correlation with CD4+ and CD8+ lymphocyte counts and alloreactive response. These findings highlight the clinical utility of immune-specific EV analysis, including immune derived EV enumeration, proteomic composition, and microRNA cargo analysis. Direct enumeration and staining of EVs from blood or plasma using advanced imaging techniques have limitations in comprehensive diagnostics. This is due to challenges such as the inability to establish a clear correlation with GVHD state when counting all EVs and the scarcity of immune-specific EVs in plasma compared to other types. A major hurdle is the need for microRNA cargo and proteomic analysis of EVs to achieve sufficient detection sensitivity, particularly for acute GVHD. MicroRNA cargo analysis requires purification of immune-derived EVs, however, existing EV sorting methods have shortcomings in effectively purifying immune specific EV subpopulations. Size/density-based methods capture all EVs but can't distinguish tissue-specific subpopulations. Affinity-based methods, like immunomagnetic sorting, offer cost-effective options but can isolate based on only one marker at a time. Combining size/density with affinity-based methods is possible but costly and space-intensive.

Ferrologix’ Digital Magnetic Sorting (DMS) technology addresses these gaps as it enables multiplexed magnetic sorting to separate biological populations based on magnetic intensity, similar to FACS sorting with dyes of different excitation/emission. Using magnetic beads of differing size and iron oxide content, DMS sorting can achieve rapid purification of EVs directly from plasma or serum targeting universal EV tetraspanins as well as immune specific markers.

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