Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine

被引：123

作者：

Oeyen, Eline ^{[1
,2
]}

Van Mol, Kurt ^{[3
]}

Baggerman, Geert ^{[1
,2
]}

Willems, Hanny ^{[1
,2
]}

Boonen, Kurt ^{[1
,2
]}

Rolfo, Christian ^{[4
,5
]}

Pauwels, Patrick ^{[6
]}

Jacobs, An ^{[1
]}

Schildermans, Karin ^{[2
]}

Cho, William C. ^{[7
]}

Mertens, Inge ^{[1
,2
]}

机构：

[1] Flemish Inst Technol Res VITO, Sustainable Hlth Dept, Mol, Belgium

[2] Univ Antwerp, Ctr Prote CFP, Antwerp, Belgium

[3] Pharmafluidics, Ghent, Belgium

[4] Antwerp Univ Hosp UZA, Phase Early Clin Trials Unit 1, Oncol Dept, Antwerp, Belgium

[5] Univ Antwerp, Ctr Oncol Res, Antwerp, Belgium

[6] Antwerp Univ Hosp UZA, Pathol Anat Dept, Edegem, Belgium

[7] Queen Elizabeth Hosp, Dept Clin Oncol, Kowloon, Hong Kong, Peoples R China

来源：

JOURNAL OF EXTRACELLULAR VESICLES | 2018年 / 7卷 / 01期

关键词：

Extracellular vesicles; small extracellular vesicles; exosomes; urine; isolation; size exclusion chromatography; asymmetrical-flow field-flow fractionation; PROTEOMIC ANALYSIS; EXOSOMES; MICROPARTICLES; MICROVESICLES; STORAGE;

D O I：

10.1080/20013078.2018.1490143

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Extracellular vesicles (EVs) have a great potential in clinical applications. However, their isolation from different bodily fluids and their characterisation are currently not optimal or standardised. Here, we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is an efficient method and provides high purity. Furthermore, we introduce asymmetrical-flow field-flow fractionation coupled with a UV detector and multi-angle light-scattering detector (AF4/UV-MALS) as a characterisation method and compare it with current methods. We demonstrate that AF4/UV-MALS is a straightforward and reproducible method for determining size, amount and purity of isolated urinary EVs.

引用

页数：12

共 43 条

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