scholarly journals Quantitative proteomic analysis of extracellular vesicle subgroups isolated by an optimized method combining polymer‐based precipitation and size exclusion chromatography

2021 ◽  
Vol 10 (6) ◽  
Author(s):  
Juan A. Martínez‐Greene ◽  
Karina Hernández‐Ortega ◽  
Ricardo Quiroz‐Baez ◽  
Osbaldo Resendis‐Antonio ◽  
Israel Pichardo‐Casas ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Proteomic Analysis ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Quantitative Proteomic Analysis ◽  
Exclusion Chromatography

Extracellular vesicle isolation methods: rising impact of size-exclusion chromatography

2019 ◽  
Vol 76 (12) ◽  
pp. 2369-2382 ◽  
Author(s):  
Marta Monguió-Tortajada ◽  
Carolina Gálvez-Montón ◽  
Antoni Bayes-Genis ◽  
Santiago Roura ◽  
Francesc E. Borràs
Keyword(s):  
Size Exclusion Chromatography ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Isolation Methods ◽  
Exclusion Chromatography

scholarly journals Size-Exclusion Chromatography as a Technique for the Investigation of Novel Extracellular Vesicles in Cancer

Cancers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3156
Author(s):  
Daniel S. K. Liu ◽  
Flora M. Upton ◽  
Eleanor Rees ◽  
Christopher Limb ◽  
Long R. Jiao ◽  
...  
Keyword(s):  
Systematic Review ◽  
Cancer Cells ◽  
Size Exclusion Chromatography ◽  
Extracellular Vesicles ◽  
Biomarker Discovery ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Isolation Method ◽  
Isolation And Purification ◽  
Exclusion Chromatography

Cancer cells release extracellular vesicles, which are a rich target for biomarker discovery and provide a promising mechanism for liquid biopsy. Size-exclusion chromatography (SEC) is an increasingly popular technique, which has been rediscovered for the purposes of extracellular vesicle (EV) isolation and purification from diverse biofluids. A systematic review was undertaken to identify all papers that described size exclusion as their primary EV isolation method in cancer research. In all, 37 papers were identified and discussed, which showcases the breadth of applications in which EVs can be utilised, from proteomics, to RNA, and through to functionality. A range of different methods are highlighted, with Sepharose-based techniques predominating. EVs isolated using SEC are able to identify cancer cells, highlight active pathways in tumourigenesis, clinically distinguish cohorts, and remain functionally active for further experiments.

scholarly journals Combination of size-exclusion chromatography and ultracentrifugation improves the proteomic profiling of plasma derived small extracellular vesicles

2020 ◽  
Author(s):  
Rui Wei ◽  
Libo Zhao ◽  
Guanyi Kong ◽  
Xiang Liu ◽  
Shengtao Zhu ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Extracellular Vesicles ◽  
Proteomic Analysis ◽  
Single Step ◽  
Size Exclusion ◽  
Systematic Evaluation ◽  
Proteomic Profiling ◽  
Exclusion Chromatography ◽  
Associated Proteins ◽  
No Gold Standard

Abstract Background: Circulating small extracellular vesicles (sEVs) and its associated proteins are of great interest in the early detection of many diseases. However, there is no gold standard for plasma sEVs isolation, especially for proteomic profiling which could be largely affected by contamination such as lipoproteins. Previous studies suggested combinations of different sEVs isolation methods could improve the purity of the isolated fractions. Nevertheless, there is no systematic evaluation of size-exclusion chromatography (SEC), ultracentrifugation (UC) and their combination in a proteomic perspective. Results: Here we exhibited that SEC+UC showed comparable recovery of sEVs with higher purity in contrast to single-step UC or SEC isolation. In our proteomic analysis, there are 992 protein species identified in the sEVs fractions isolated by SEC+UC, much more than the sEVs fractions isolated by UC (453) or SEC (682) alone. As compared to Vesiclepedia and Exocarta databases, SEC+UC kept 584 previously identified sEV-associated proteins and 360 other proteins. Furthermore, detailed analysis suggested that sEV-associated proteins, such as CD9, CD81 and ITGB1, showed the better protein rank in SEC+UC group than UC group and SEC group. Lipoproteins, the most common contamination in sEVs proteomic analysis, along with other free-floating proteins in the plasma, were largely removed in SEC+UC. Conclusions: We suggested that combining SEC with UC could significantly improve the performance of mass-spectrum (MS)-based proteomic profiling in analyzing plasma-derived sEVs.

scholarly journals Mass-Spectrometry Based Proteome Comparison of Extracellular Vesicle Isolation Methods: Comparison of ME-kit, Size-Exclusion Chromatography, and High-Speed Centrifugation

Biomedicines ◽  
2020 ◽  
Vol 8 (8) ◽  
pp. 246 ◽  
Author(s):  
Anders Askeland ◽  
Anne Borup ◽  
Ole Østergaard ◽  
Jesper V. Olsen ◽  
Sigrid M. Lund ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Size Exclusion Chromatography ◽  
High Speed ◽  
Proteome Analysis ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Label Free ◽  
Protein Biomarkers ◽  
Transmission Electron ◽  
Exclusion Chromatography

Extracellular vesicles (EVs) are small membrane-enclosed particles released by cells under various conditions specific to cells’ biological states. Hence, mass-spectrometry (MS) based proteome analysis of EVs in plasma has gained much attention as a method to discover novel protein biomarkers. MS analysis of EVs in plasma is challenging and EV isolation is usually necessary. Therefore, we compared differences in abundance, subtypes, and contamination for EVs isolated by high-speed centrifugation, size exclusion chromatography (SEC), and peptide-affinity precipitation (PAP/ME kit) for subsequent MS-based proteome analysis. Successful EV isolation was evaluated by nanoparticle-tracking analysis, immunoblotting, and transmission electron microscopy, while EV abundance, EV subtypes, and contamination was evaluated by label-free tandem MS. High-speed centrifugation and SEC isolates showed high EV abundance at the expense of contamination by non-EV proteins and lipoproteins, respectively. These two methods also resulted in EVs of a similar type, however, with smaller EVs in SEC isolates. PAP isolates had a relatively low EV abundance and high contamination. We consider high-speed centrifugation and SEC suitable as EV isolation for MS biomarker studies, where the choice between the two should depend on the scientific questions and whether the focus is on larger or smaller EVs or a combination of both.

scholarly journals Efficient extracellular vesicle isolation by combining cell media modifications, ultrafiltration, and size-exclusion chromatography

PLoS ONE ◽  
2018 ◽  
Vol 13 (9) ◽  
pp. e0204276 ◽  
Author(s):  
Eduarda M. Guerreiro ◽  
Beate Vestad ◽  
Lilly Alice Steffensen ◽  
Hans Christian D. Aass ◽  
Muhammad Saeed ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Exclusion Chromatography

Extracellular‐Vesicle Isolation from Different Biological Fluids by Size‐Exclusion Chromatography

2019 ◽  
Vol 49 (1) ◽  
pp. e82 ◽  
Author(s):  
Marta Monguió‐Tortajada ◽  
Miriam Morón‐Font ◽  
Ana Gámez‐Valero ◽  
Laura Carreras‐Planella ◽  
Francesc E. Borràs ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Biological Fluids ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Exclusion Chromatography
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