Vedrana Krušić Alić (Author), Mladenka Malenica (Author), Maša Biberić (Author), Siniša Zrna (Author), Lara Valenčić (Author), Aleksandar Šuput (Author), Lada Kalagac Fabris (Author), Karmen Wechtersbach (Author), Nika Kojc (Author), Mario Kurtjak (Author)

Abstract

Extracellular vesicles (EVs) are a versatile group of cell-secreted membranous nanoparticles present in body fluids. They have an exceptional diagnostic potential due to their molecular content matching the originating cells and accessibility from body fluids. However, methods for EV isolation are still in development, with size exclusion chromatography (SEC) emerging as a preferred method. Here we compared four types of SEC to isolate EVs from the CSF of patients with severe traumatic brain injury. A pool of nine CSF samples was separated by SEC columns packed with Sepharose CL-6B, Sephacryl S-400 or Superose 6PG and a ready-to-use qEV10/70 nm column. A total of 46 fractions were collected and analysed by slot-blot followed by Ponceau staining. Immunodetection was performed for albumin, EV markers CD9, CD81, and lipoprotein markers ApoE and ApoAI. The size and concentration of nanoparticles in fractions were determined by tunable resistive pulse sensing and EVs were visualised by transmission electron microscopy. We show that all four SEC techniques enabled separation of CSF into nanoparticle- and free protein-enriched fractions. Sepharose CL-6B resulted in a significantly higher number of separated EVs while lipoproteins were eluted together with free proteins. Our data indicate that Sepharose CL-6B is suitable for isolation of EVs from CSF and their separation from lipoproteins.

Keywords

extracellular vesicles;cerebrospinal fluid;traumatic brain injury;size exclusion chromatography;CD9 protein;CD81 protein;apolipoproteins;

Data

Language: English
Year of publishing:
Typology: 1.01 - Original Scientific Article
Organization: UL MF - Faculty of Medicine
UDC: 577.2
COBISS: 119230211 Link will open in a new window
ISSN: 2227-9059
Views: 150
Downloads: 50
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Other data

Secondary language: Slovenian
Secondary keywords: zunajcelični vezikli;cerebrospinalna tekočina;travmatska poškodba možganov;velikostno izločitvena kromatografija;protein CD9;protein CD81;apolipoproteini;
Type (COBISS): Article
Pages: 1-15 str.
Volume: ǂVol. ǂ10
Issue: ǂiss. ǂ4
Chronology: 2022
DOI: 10.3390/biomedicines10040785
ID: 15721005