Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry

Leonie de Rond; Edwin van der Pol; Chi M. Hau; Zoltan Varga; Auguste Sturk; Ton G. van Leeuwen; Rienk Nieuwland; Frank A. W. Coumans

doi:https://doi.org/10.1373/clinchem.2017.278978

Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry

Leonie de Rond, Edwin van der Pol, Chi M. Hau, Zoltan Varga, Auguste Sturk, Ton G. van Leeuwen, Rienk Nieuwland, Frank A. W. Coumans

Research output: Contribution to journal › Article › Academic › peer-review

67 Citations (Scopus)

Abstract

Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. +)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. +EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used

Original language	English
Pages (from-to)	680-689
Journal	Clinical Chemistry
Volume	64
Issue number	4
DOIs	https://doi.org/10.1373/clinchem.2017.278978
Publication status	Published - 2018

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https://doi.org/10.1373/clinchem.2017.278978

Cite this

@article{1d80ba245b464e6da0c0e6f10d0f3d20,

title = "Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry",

abstract = "Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. +)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. +EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used",

author = "{de Rond}, Leonie and {van der Pol}, Edwin and Hau, {Chi M.} and Zoltan Varga and Auguste Sturk and {van Leeuwen}, {Ton G.} and Rienk Nieuwland and Coumans, {Frank A. W.}",

year = "2018",

doi = "https://doi.org/10.1373/clinchem.2017.278978",

language = "English",

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T1 - Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry

AU - de Rond, Leonie

AU - van der Pol, Edwin

AU - Hau, Chi M.

AU - Varga, Zoltan

AU - Sturk, Auguste

AU - van Leeuwen, Ton G.

AU - Nieuwland, Rienk

AU - Coumans, Frank A. W.

PY - 2018

Y1 - 2018

N2 - Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. +)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. +EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used

AB - Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. +)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. +EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used

U2 - https://doi.org/10.1373/clinchem.2017.278978

DO - https://doi.org/10.1373/clinchem.2017.278978

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SN - 0009-9147

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