Research Project: Exosome Characterization Platform for Early Detection of Breast Cancer
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Contributors
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EC.00165
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EV-Lev: extracellular vesicle isolation from human plasma using microfluidic magnetic levitation device
(Royal Society of Chemistry, 2025) Aygün, Uğur; Aygün, Uğur; Yaman, Sena; Devoe, Tessa; Parlatan, Ugur; Bobbili, Madhusudhan Reddy; Karim, Asma H.; Grillari, Johannes; Durmus, Naside Gozde; Department of Electrical and Electronics Engineering; No; College of Engineering
Biological nanomaterials have unique magnetic and density characteristics that can be employed to isolate them into subpopulations. Extracellular nanovesicles (EVs) are crucial for cellular communication; however, their isolation poses significant challenges due to their diverse sizes and compositions. We present EV-Lev, a microfluidic magnetic levitation technique for high-throughput, selective isolation of small EVs (<200 nm) from human plasma. EV-Lev overcomes the challenges posed by the subtle buoyancy characteristics of EVs, whose small size and varied densities complicate traditional magnetic levitation techniques. It employs antibody-coated polymer beads of varying densities, integrating immuno-affinity and microfluidics to isolate EVs from sub-milliliter plasma volumes efficiently. It facilitates rapid, simultaneous sorting of EV subpopulations based on surface markers, such as CD9, CD63, and CD81, achieving high yield and purity. Subsequent size and morphology analyses confirmed that the isolated EVs maintain their structural integrity. EV-Lev could help uncover the cargo and function of EV subpopulations associated with multiple diseases including cancer, infectious diseases and help to discover potential biomarkers in small volume samples, while offering a portable, cost-effective, and straightforward assay scheme.
Nanoscale detection of EpCAM-positive extracellular vesicles using magnetic beads and interferometric scattering microscopy
(2025) Aygün, Uğur; Aygün, Uğur; Yaman, Sena; Parlatan, Ugur; Durmus, Naside Gozde; Department of Electrical and Electronics Engineering; No; College of Engineering
Extracellular vesicles (EVs) are nanoscale biomarkers that reflect the molecular state of their cell of origin. Epithelial cell adhesion molecule (EpCAM) is frequently enriched in tumor-derived EVs, offering diagnostic and monitoring potential for epithelial cancers. We developed a label-free detection method combining magnetic bead–based molecular targeting with interferometric scattering microscopy (iSCAT) for single-particle visualization of EpCAM-positive EVs. Streptavidin-coated magnetic nanoparticles (50 nm) were conjugated with biotinylated anti-EpCAM antibodies and incubated with EVs from breast cancer (MCF-7, high EpCAM) and murine colon adenocarcinoma (MC38, low EpCAM) cell lines. Nanoparticle tracking analysis confirmed bead functionalization, and Western blotting verified differential EpCAM expression. iSCAT imaging on thin-film substrates revealed distinct scattering patterns: bare beads showed uniform profiles, high EpCAM EV–bead complexes exhibited elongated morphologies, and low EpCAM samples showed minimal change. The method requires <1 µL of sample, avoids labeling artifacts, and enables real-time, high-contrast imaging. This adaptable platform can be tailored to other EV subpopulations, providing a sensitive, label-free approach for targeted EV detection in research and clinical diagnostics.
