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Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations

AIM: For isolation of exosomes, differential ultracentrifugation and an isolation kit from a major vendor were compared. MATERIALS & METHODS: ‘Case study’ exosomes isolated from patient-derived cells from glioblastoma multiforme and a breast cancer cell line were analyzed. RESULTS: Transmission...

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Autores principales: Skottvoll, Frøydis Sved, Berg, Henriette Engen, Bjørseth, Kamilla, Lund, Kaja, Roos, Norbert, Bekhradnia, Sara, Thiede, Bernd, Sandberg, Cecilie, Vik-Mo, Einar Osland, Roberg-Larsen, Hanne, Nyström, Bo, Lundanes, Elsa, Wilson, Steven Ray
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6331754/
https://www.ncbi.nlm.nih.gov/pubmed/30652024
http://dx.doi.org/10.4155/fsoa-2018-0088
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author Skottvoll, Frøydis Sved
Berg, Henriette Engen
Bjørseth, Kamilla
Lund, Kaja
Roos, Norbert
Bekhradnia, Sara
Thiede, Bernd
Sandberg, Cecilie
Vik-Mo, Einar Osland
Roberg-Larsen, Hanne
Nyström, Bo
Lundanes, Elsa
Wilson, Steven Ray
author_facet Skottvoll, Frøydis Sved
Berg, Henriette Engen
Bjørseth, Kamilla
Lund, Kaja
Roos, Norbert
Bekhradnia, Sara
Thiede, Bernd
Sandberg, Cecilie
Vik-Mo, Einar Osland
Roberg-Larsen, Hanne
Nyström, Bo
Lundanes, Elsa
Wilson, Steven Ray
author_sort Skottvoll, Frøydis Sved
collection PubMed
description AIM: For isolation of exosomes, differential ultracentrifugation and an isolation kit from a major vendor were compared. MATERIALS & METHODS: ‘Case study’ exosomes isolated from patient-derived cells from glioblastoma multiforme and a breast cancer cell line were analyzed. RESULTS: Transmission electron microscopy, dynamic light scattering, western blotting, and so forth, revealed comparable performance. Potential protein biomarkers for both diseases were also identified in the isolates using nanoLC–MS. Western blotting and nanoLC–MS also revealed negative exosome markers regarding both isolation approaches. CONCLUSION: The two isolation methods had an overall similar performance, but we hesitate to use the term ‘exosome isolation’ as impurities may be present with both isolation methods. NanoLC–MS can detect disease biomarkers in exosomes and is useful for critical assessment of exosome enrichment procedures.
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spelling pubmed-63317542019-01-16 Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations Skottvoll, Frøydis Sved Berg, Henriette Engen Bjørseth, Kamilla Lund, Kaja Roos, Norbert Bekhradnia, Sara Thiede, Bernd Sandberg, Cecilie Vik-Mo, Einar Osland Roberg-Larsen, Hanne Nyström, Bo Lundanes, Elsa Wilson, Steven Ray Future Sci OA Research Article AIM: For isolation of exosomes, differential ultracentrifugation and an isolation kit from a major vendor were compared. MATERIALS & METHODS: ‘Case study’ exosomes isolated from patient-derived cells from glioblastoma multiforme and a breast cancer cell line were analyzed. RESULTS: Transmission electron microscopy, dynamic light scattering, western blotting, and so forth, revealed comparable performance. Potential protein biomarkers for both diseases were also identified in the isolates using nanoLC–MS. Western blotting and nanoLC–MS also revealed negative exosome markers regarding both isolation approaches. CONCLUSION: The two isolation methods had an overall similar performance, but we hesitate to use the term ‘exosome isolation’ as impurities may be present with both isolation methods. NanoLC–MS can detect disease biomarkers in exosomes and is useful for critical assessment of exosome enrichment procedures. Future Science Ltd 2018-11-09 /pmc/articles/PMC6331754/ /pubmed/30652024 http://dx.doi.org/10.4155/fsoa-2018-0088 Text en © 2018 The Authors This work is licensed under a Creative Commons Attribution 4.0 License (http://creativecommons.org/licenses/by/4.0/)
spellingShingle Research Article
Skottvoll, Frøydis Sved
Berg, Henriette Engen
Bjørseth, Kamilla
Lund, Kaja
Roos, Norbert
Bekhradnia, Sara
Thiede, Bernd
Sandberg, Cecilie
Vik-Mo, Einar Osland
Roberg-Larsen, Hanne
Nyström, Bo
Lundanes, Elsa
Wilson, Steven Ray
Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations
title Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations
title_full Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations
title_fullStr Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations
title_full_unstemmed Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations
title_short Ultracentrifugation versus kit exosome isolation: nanoLC–MS and other tools reveal similar performance biomarkers, but also contaminations
title_sort ultracentrifugation versus kit exosome isolation: nanolc–ms and other tools reveal similar performance biomarkers, but also contaminations
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6331754/
https://www.ncbi.nlm.nih.gov/pubmed/30652024
http://dx.doi.org/10.4155/fsoa-2018-0088
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