Naslov (eng)

Influence of SEC to resistance of EVs to purity control treatments and stability in different handling/storage conditions

Autor

Gruden-Movsesijan, Alisa
Glamočlija, Sofija
Kosanović, Maja
Bekić, Marina
Tomić, Sergej

Publisher

Opis (eng)

ABSTRACT Introduction: To utilize diagnostic/therapeutic potential of extracellular vesicles (EVs), their integrity should be kept throughout the separation, purification and storage process. However, some techniques, such as size exclusion chromatography (SEC), can affect the corona of EVs and influence both diagnostic/therapeutic molecules and stability of EVs. Therefore, the aim of this work was to investigate whether SEC affects EVs’ resistance to purity control treatments and stability in different handling/storage conditions. Materials & Methods: We separated EVs by ultracentrifugation (UC) on 17000xg (T17) and 100.000xg (T100) and subjected half of obtained EVs to SEC (Sepharose 2B CL, 11 x 1.2 cm column). In each fraction, the number of particles was determined by NTA, protein concentration by BCA and EVs’ marker profile by dot-blot. Fractions containing peak of EVs were pooled. EVs before and after SEC were incubated with different detergents, buffers and were subjected to different temperatures after which the median size and concentration were determined by NTA. Tripsin was used for direct removal of corona.

Jezik

engleski

Datum

2023

Licenca

© All rights reserved

Predmet

extracellular vesicles (EVs), exclusion chromatography (SEC)

Deo kolekcije (1)

o:25140

Radovi Instituta za primenu nuklearne energije - INEP