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

Povzetek

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.

Ključne besede

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

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 1.01 - Izvirni znanstveni članek
Organizacija: UL MF - Medicinska fakulteta
UDK: 577.2
COBISS: 119230211 Povezava se bo odprla v novem oknu
ISSN: 2227-9059
Št. ogledov: 150
Št. prenosov: 50
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Slovenski jezik
Sekundarne ključne besede: zunajcelični vezikli;cerebrospinalna tekočina;travmatska poškodba možganov;velikostno izločitvena kromatografija;protein CD9;protein CD81;apolipoproteini;
Vrsta dela (COBISS): Članek v reviji
Strani: 1-15 str.
Letnik: ǂVol. ǂ10
Zvezek: ǂiss. ǂ4
Čas izdaje: 2022
DOI: 10.3390/biomedicines10040785
ID: 15721005