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Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination

Given the emerging diagnostic utility of extracellular vesicles (EVs), it is important to account for non-EV contaminants. Lipoprotein present in EV-enriched isolates may inflate particle counts and decrease sensitivity to biomarkers of interest, skewing chemical analyses and perpetuating downstream...

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Autores principales: Koster, Hanna J., Rojalin, Tatu, Powell, Alyssa, Pham, Dina, Mizenko, Rachel R., Birkeland, Andrew C., Carney, Randy P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8447870/
https://www.ncbi.nlm.nih.gov/pubmed/34473170
http://dx.doi.org/10.1039/d1nr03334d
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author Koster, Hanna J.
Rojalin, Tatu
Powell, Alyssa
Pham, Dina
Mizenko, Rachel R.
Birkeland, Andrew C.
Carney, Randy P.
author_facet Koster, Hanna J.
Rojalin, Tatu
Powell, Alyssa
Pham, Dina
Mizenko, Rachel R.
Birkeland, Andrew C.
Carney, Randy P.
author_sort Koster, Hanna J.
collection PubMed
description Given the emerging diagnostic utility of extracellular vesicles (EVs), it is important to account for non-EV contaminants. Lipoprotein present in EV-enriched isolates may inflate particle counts and decrease sensitivity to biomarkers of interest, skewing chemical analyses and perpetuating downstream issues in labeling or functional analysis. Using label free surface enhanced Raman scattering (SERS), we confirm that three common EV isolation methods (differential ultracentrifugation, density gradient ultracentrifugation, and size exclusion chromatography) yield variable lipoprotein content. We demonstrate that a dual-isolation method is necessary to isolate EVs from the major classes of lipoprotein. However, combining SERS analysis with machine learning assisted classification, we show that the disease state is the main driver of distinction between EV samples, and largely unaffected by choice of isolation. Ultimately, this study describes a convenient SERS assay to retain accurate diagnostic information from clinical samples by overcoming differences in lipoprotein contamination according to isolation method.
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spelling pubmed-84478702021-10-18 Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination Koster, Hanna J. Rojalin, Tatu Powell, Alyssa Pham, Dina Mizenko, Rachel R. Birkeland, Andrew C. Carney, Randy P. Nanoscale Chemistry Given the emerging diagnostic utility of extracellular vesicles (EVs), it is important to account for non-EV contaminants. Lipoprotein present in EV-enriched isolates may inflate particle counts and decrease sensitivity to biomarkers of interest, skewing chemical analyses and perpetuating downstream issues in labeling or functional analysis. Using label free surface enhanced Raman scattering (SERS), we confirm that three common EV isolation methods (differential ultracentrifugation, density gradient ultracentrifugation, and size exclusion chromatography) yield variable lipoprotein content. We demonstrate that a dual-isolation method is necessary to isolate EVs from the major classes of lipoprotein. However, combining SERS analysis with machine learning assisted classification, we show that the disease state is the main driver of distinction between EV samples, and largely unaffected by choice of isolation. Ultimately, this study describes a convenient SERS assay to retain accurate diagnostic information from clinical samples by overcoming differences in lipoprotein contamination according to isolation method. The Royal Society of Chemistry 2021-08-26 /pmc/articles/PMC8447870/ /pubmed/34473170 http://dx.doi.org/10.1039/d1nr03334d Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Koster, Hanna J.
Rojalin, Tatu
Powell, Alyssa
Pham, Dina
Mizenko, Rachel R.
Birkeland, Andrew C.
Carney, Randy P.
Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
title Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
title_full Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
title_fullStr Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
title_full_unstemmed Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
title_short Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
title_sort surface enhanced raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8447870/
https://www.ncbi.nlm.nih.gov/pubmed/34473170
http://dx.doi.org/10.1039/d1nr03334d
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