UFLC-Derived CSF Extracellular Vesicle Origin and Proteome.
Thompson AG., Gray E., Mager I., Fischer R., Thézénas M-L., Charles PD., Talbot K., El Andaloussi S., Kessler BM., Wood M., Turner MR.
Cerebrospinal fluid (CSF) extracellular vesicles (EVs) show promise as a source of neurological disease biomarkers, although their precise origin is poorly understood. Current extraction techniques produce disappointing yield and purity. This study describes the application of ultrafiltration LC (UFLC) to CSF-EVs, compared with ultracentrifugation (UC), and explores CSF-EV origin. EVs are extracted from human CSF by UC and UFLC and characterized using nanoparticle tracking analysis, electron microscopy, and immunoblotting. EV and CSF proteomes are analyzed by LC-MS/MS. UFLC-isolated particles have size, morphology, and marker expression characteristic of EVs. UFLC provides greater EV yield (UFLC 7.90 × 108 ± SD 1.31 × 108 EVs mL-1 CSF, UC 1.06 × 108 ± 0.57 × 108 p