Journal: Neurology® Neuroimmunology & Neuroinflammation
Article Title: Extracellular Vesicle Marker Changes Associated With Disease Activity in Relapsing-Remitting Multiple Sclerosis
doi: 10.1212/NXI.0000000000200404
Figure Lengend Snippet: (A) Ultrasensitive detection of neurofilament light chain (NfL) and glial fibrillary acidic protein (GFAP) by single-molecule array (SIMOA) in plasma samples. (B) EVs were analyzed using the single-particle interferometric resonance imaging sensor (SP-IRIS, ExoView, Unchained Labs) after immunocapture with CD9, CD81, and CD63 antibodies directly in plasma. (C) EV isolation with size-exclusion chromatography (SmartSEC, System Biosciences) followed by the characterization of isolated EVs by Western blotting (WB), nanoparticle-tracking analysis (NTA), and cryo-electron microscopy (cryo-EM). Extended EV-specific marker analysis by bead-based flow cytometry (MACSPlex EV kit, Miltenyi) after size-exclusion chromatography-based EV isolation. The levels of 37 EV surface markers plus 2 isotype controls (MIgG, human IgG1 isotype control antibody [REA293]) were simultaneously monitored. Each type of EV capture bead was labeled with 1 of the 39 antibodies. Bound EVs were detected with a cocktail of APC-labeled CD9, CD81, and CD63 antibodies. Illustrations created in BioRender Falk, V. (2025) BioRender.com/uqfyfpg and Adobe Photoshop (v. 2024). EV = extracellular vesicle.
Article Snippet: Single-particle interferometric resonance imaging sensor (SP-IRIS) with ExoView (Unchained Labs, Pleasanton, CA) enables size and density measurement of EVs exhibiting membrane-bound tetraspanins CD9, CD63, and CD81 in crude plasma, without previous EV isolation.
Techniques: Clinical Proteomics, Single Particle, Imaging, Isolation, Size-exclusion Chromatography, Western Blot, Cryo-Electron Microscopy, Cryo-EM Sample Prep, Marker, Flow Cytometry, Control, Labeling