Naslov (eng)

Urinary extracellular vesicles as a readily available biomarker source: a simplified stratification method

Autor

Filipović, Lidija
Spasojević Savković, Milica
Prodanović, Radivoje
Matijašević Joković, Suzana
Stevanović, Sanja
de Marco, Ario
Kosanović, Maja
Brajušković, Goran
Popović, Milica

Publisher

Opis (eng)

ABSTRACT Urine, a common source of biological markers in biomedical research and clinical diagnosis, has recently generated a new wave of interest. It has recently become a focus of study due to the presence of its content of extracellular vesicles (EVs). These uEVs have been found to reflectphysiological and pathological conditions in kidney, urothelial, and prostate tissue and can illustrate further molecular processes, leading to a rapid expansion of research in this field In this work,we present the advantages of an immunoaffinity-based method for uEVs’ isolation with respect to the gold standard purification approach performed by differential ultracentrifugation [in terms of purity and antigen presence. The immunoaffinity method was made feasible by combining specific antibodies with a functionalized polymethacrylate polymer. Flow cytometry indicated a significant fluorescence shift, validating the presence of the markers (CD9, CD63, CD81) and confirming the effectiveness of the isolation method. Microscopy evaluations have shown that the morphology of the vesicles remained intact and corresponded to the expected shapes and dimensions of uEVs. The described protocol is inexpensive, fast, easy to process, has good reproducibility, and can be applied to further biological samples.

Jezik

engleski

Datum

2024

Licenca

Ovo delo je licencirano pod uslovima licence
Creative Commons CC BY 4.0 - Creative Commons Autorstvo 4.0 International License.

http://creativecommons.org/licenses/by/4.0/legalcode

Predmet

extracellular vesicles; nanobodies; immunoaffinity chromatography; ultracentrifugation

Deo kolekcije (1)

o:25140

Radovi Instituta za primenu nuklearne energije - INEP