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. 2022 Mar 27;10(4):785. doi: 10.3390/biomedicines10040785

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Nanoparticles isolated by Sepharose CL-6B contain CD9+ and CD81+ EVs. (a) CSF-pool from severe traumatic brain injury patients was separated by a gravity-flow column packed with sepharose CL-6B. Consecutively collected fractions indicated by numbers together with CSF-pool as positive control (PC) and PBS (mobile phase in SEC) as negative control (NC) were applied to nitrocellulose membrane by slot blot and analysed by immunodetection of indicated proteins. Shown are representative immunoblots after chemiluminescence. The chemiluminescence signal after immunodetection on slot blots for indicated proteins was quantified and normalised to the PC signal. Shown are relative protein abundances obtained by the quantification of representative immunoblots (b). Selected fractions were analysed by western blot after protein separation by SDS-PAGE. (c) Transmission electron microscopy micrograph showing round shaped extracellular vesicles measuring 100–130 nm in diameter and indicated by arrows (A). Higher magnification of electron micrograph showing extracellular vesicle with double-layer membrane measuring 175 nm (B).