Extracellular vesicles from human cerebrospinal fluid are effectively separated by Sepharose CL-6B—comparison of four gravity-flow size exclusion chromatography methods

Krušić Alić, Vedrana (avtor), Wechtersbach, Karmen (avtor), Kojc, Nika (avtor), Grabušić, Kristina (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: extracellular vesicles, cerebrospinal fluid, traumatic brain injury, size exclusion chromatography, CD9 protein, CD81 protein, apolipoproteins
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: MF - Medicinska fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2022
• Št. strani: 15 str.
• Številčenje: Vol. 10, iss. 4, art. 785
• PID: 20.500.12556/RUL-137627
• ISSN pri članku: 2227-9059
• DOI: 10.3390/biomedicines10040785
• COBISS.SI-ID: 119230211
• Datum objave v RUL: 23.06.2022

Gradivo je del revije

• Naslov: Biomedicines
• Skrajšan naslov: Biomedicines
• Založnik: MDPI AG
• ISSN: 2227-9059
• COBISS.SI-ID: 523006745

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: zunajcelični vezikli, cerebrospinalna tekočina, travmatska poškodba možganov, velikostno izločitvena kromatografija, protein CD9, protein CD81, apolipoproteini