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Extracellular vesicles from human cerebrospinal fluid are effectively separated by Sepharose CL-6B—comparison of four gravity-flow size exclusion chromatography methods
ID
Krušić Alić, Vedrana
(
Avtor
),
ID
Wechtersbach, Karmen
(
Avtor
),
ID
Kojc, Nika
(
Avtor
),
ID
Grabušić, Kristina
(
Avtor
), et al.
PDF - Predstavitvena datoteka,
prenos
(1,62 MB)
MD5: 181E55D03396290F1184333E8DF910BE
URL - Izvorni URL, za dostop obiščite
https://www.mdpi.com/2227-9059/10/4/785
Galerija slik
Izvleček
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
Število ogledov:
607
Število prenosov:
107
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Objavi na:
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
Projekti
Financer:
HRZZ - Croatian Science Foundation
Številka projekta:
IP-2019-04-1511
Financer:
Drugi - Drug financer ali več financerjev
Program financ.:
University of Rijeka
Številka projekta:
uniri-biomed-18-5
Financer:
Drugi - Drug financer ali več financerjev
Program financ.:
University of Rijeka
Številka projekta:
uniri-biomed-18-279
Financer:
ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Številka projekta:
P3-0054
Naslov:
Patologija in molekularna genetika
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