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; Malenica, Mladenka; Biberić, Maša; Zrna, Siniša; Valenčić, Lara; Šuput, Aleksandar; Kalagac Fabris, Lada; Wechtersbach, Karmen; Kojc, Nika; Kurtjak, Mario; Kučić, Natalia; Grabušić, Kristina

doi:10.3390/biomedicines10040785

Pregled bibliografske jedinice broj: 1211858

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; Malenica, Mladenka; Biberić, Maša; Zrna, Siniša; Valenčić, Lara; Šuput, Aleksandar; Kalagac Fabris, Lada; Wechtersbach, Karmen; Kojc, Nika; Kurtjak, Mario et al.

Extracellular Vesicles from Human Cerebrospinal Fluid Are Effectively Separated by Sepharose CL-6B —Comparison of Four Gravity-Flow Size Exclusion Chromatography Methods // Biomedicines, 10 (2022), 4; 785, 10 doi:10.3390/biomedicines10040785 (međunarodna recenzija, članak, znanstveni)

CROSBI ID: 1211858 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Extracellular Vesicles from Human Cerebrospinal Fluid Are Effectively Separated by Sepharose CL-6B —Comparison of Four Gravity-Flow Size Exclusion Chromatography Methods

Autori
Krušić Alić, Vedrana ; Malenica, Mladenka ; Biberić, Maša ; Zrna, Siniša ; Valenčić, Lara ; Šuput, Aleksandar ; Kalagac Fabris, Lada ; Wechtersbach, Karmen ; Kojc, Nika ; Kurtjak, Mario ; Kučić, Natalia ; Grabušić, Kristina

Izvornik
Biomedicines (2227-9059) 10 (2022), 4; 785, 10

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

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

Sažetak
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.

Izvorni jezik
Engleski

POVEZANOST RADA

Projekti:
HRZZ-IP-2019-04-1511 - Identifikacija cirkulirajućih biomarkera neurološkog oporavka u bolesnika s ozljedom mozga (CiMozak / CirBrain) (Grabušić, Kristina, HRZZ - 2019-04) ( CroRIS)
--uniri-biomed-18-5 - Izvanstanične vezikule kao klinički markeri neuroregeneracije nakon teške ozljede mozga (Grabušić, Kristina; Grabušić, Kristina) ( CroRIS)

Profili:

Natalia Kučić (autor)