Quality Assessment and Comparison of Plasma-Derived Extracellular Vesicles Separated by Three Commercial Kits for Prostate Cancer Diagnosis.

Pang, B; Zhu, Y; Ni, J; Ruan, J; Thompson, J; Malouf, D; Bucci, J; Graham, P; Li, Y

Quality Assessment and Comparison of Plasma-Derived Extracellular Vesicles Separated by Three Commercial Kits for Prostate Cancer Diagnosis.

Pang, B Zhu, Y

Ni, J Ruan, J Thompson, J Malouf, D Bucci, J Graham, P Li, Y

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Publisher:: DOVE MEDICAL PRESS LTD
Publication Type:: Journal Article
Citation:: International journal of nanomedicine, 2020, 15, pp. 10241-10256
Issue Date:: 2020-01

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Field	Value	Language
dc.contributor.author	Pang, B
dc.contributor.author	Zhu, Y https://orcid.org/0000-0002-8840-176X
dc.contributor.author	Ni, J
dc.contributor.author	Ruan, J
dc.contributor.author	Thompson, J
dc.contributor.author	Malouf, D
dc.contributor.author	Bucci, J
dc.contributor.author	Graham, P
dc.contributor.author	Li, Y
dc.date.accessioned	2021-03-24T22:20:03Z
dc.date.available	2020-11-07
dc.date.available	2021-03-24T22:20:03Z
dc.date.issued	2020-01
dc.identifier.citation	International journal of nanomedicine, 2020, 15, pp. 10241-10256
dc.identifier.issn	1176-9114
dc.identifier.issn	1178-2013
dc.identifier.uri	http://hdl.handle.net/10453/147512
dc.description.abstract	<h4>Introduction</h4>Current standard biomarkers in clinic are not specific enough for prostate cancer (PCa) diagnosis. Extracellular vesicles (EVs) are nano-scale vesicles released by most mammalian cells. EVs are promising biomarker source for PCa liquid biopsy due to its minimal invasive approach, rich information and improved accuracy compared to the clinical standard prostate-specific antigen (PSA). However, current EV separation methods cannot separate pure EVs and the quality characteristics from these methods remain largely unknown. In this study, we evaluated the quality characteristics of human plasma-derived EVs by comparing three clinical suitable separation kits.<h4>Methods</h4>We combined EV separation by commercial kits with magnetic beads capture and flow cytometry analysis, and compared three kits including ExoQuick Ultra based on precipitation and qEV35 and qEV70 based on size exclusion chromatography (SEC).<h4>Results</h4>Our results indicated that two SEC kits provided higher EV purity and lower protein contamination compared to ExoQuick Ultra precipitation and that qEV35 demonstrated a higher EV yield but lower EV purity compared to qEV70. Particle number correlated very well particularly with CD9/81/63 positive EVs for all three kits, which confirms that particle number can be used as the estimate for EV amount. At last, we found that several EV metrics including total EVs and PSA-specific EVs could not differentiate PCa patients from health controls.<h4>Conclusion</h4>We provided a systematic workflow for the comparison of three separation kits as well as a general analysis process in clinical laboratories for EV-based cancer diagnosis. Better EV-associated cancer biomarkers need to be explored in the future study with a larger cohort.
dc.format	Electronic-eCollection
dc.language	eng
dc.publisher	DOVE MEDICAL PRESS LTD
dc.relation.ispartof	International journal of nanomedicine
dc.relation.isbasedon	10.2147/ijn.s283106
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	0601 Biochemistry and Cell Biology, 1007 Nanotechnology, 1115 Pharmacology and Pharmaceutical Sciences
dc.subject.classification	Nanoscience & Nanotechnology
dc.subject.mesh	Plasma
dc.subject.mesh	Humans
dc.subject.mesh	Prostatic Neoplasms
dc.subject.mesh	Chromatography, Gel
dc.subject.mesh	Immunomagnetic Separation
dc.subject.mesh	Male
dc.subject.mesh	Extracellular Vesicles
dc.subject.mesh	Chromatography, Gel
dc.subject.mesh	Extracellular Vesicles
dc.subject.mesh	Humans
dc.subject.mesh	Immunomagnetic Separation
dc.subject.mesh	Male
dc.subject.mesh	Plasma
dc.subject.mesh	Prostatic Neoplasms
dc.title	Quality Assessment and Comparison of Plasma-Derived Extracellular Vesicles Separated by Three Commercial Kits for Prostate Cancer Diagnosis.
dc.type	Journal Article
utslib.citation.volume	15
utslib.location.activity	New Zealand
utslib.for	0601 Biochemistry and Cell Biology
utslib.for	1007 Nanotechnology
utslib.for	1115 Pharmacology and Pharmaceutical Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
utslib.copyright.status	open_access	*
dc.date.updated	2021-03-24T22:19:51Z
pubs.publication-status	Published
pubs.volume	15

Abstract:

Introduction

Current standard biomarkers in clinic are not specific enough for prostate cancer (PCa) diagnosis. Extracellular vesicles (EVs) are nano-scale vesicles released by most mammalian cells. EVs are promising biomarker source for PCa liquid biopsy due to its minimal invasive approach, rich information and improved accuracy compared to the clinical standard prostate-specific antigen (PSA). However, current EV separation methods cannot separate pure EVs and the quality characteristics from these methods remain largely unknown. In this study, we evaluated the quality characteristics of human plasma-derived EVs by comparing three clinical suitable separation kits.

Methods

We combined EV separation by commercial kits with magnetic beads capture and flow cytometry analysis, and compared three kits including ExoQuick Ultra based on precipitation and qEV35 and qEV70 based on size exclusion chromatography (SEC).

Results

Our results indicated that two SEC kits provided higher EV purity and lower protein contamination compared to ExoQuick Ultra precipitation and that qEV35 demonstrated a higher EV yield but lower EV purity compared to qEV70. Particle number correlated very well particularly with CD9/81/63 positive EVs for all three kits, which confirms that particle number can be used as the estimate for EV amount. At last, we found that several EV metrics including total EVs and PSA-specific EVs could not differentiate PCa patients from health controls.

Conclusion

We provided a systematic workflow for the comparison of three separation kits as well as a general analysis process in clinical laboratories for EV-based cancer diagnosis. Better EV-associated cancer biomarkers need to be explored in the future study with a larger cohort.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/147512