Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing

Lane, Rebecca E., Korbie, Darren, Anderson, Will, Vaidyanathan, Ramanathan and Trau, Matt (2015) Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing. Scientific Reports, 5 7639.1-7639.7. doi:10.1038/srep07639


Author Lane, Rebecca E.
Korbie, Darren
Anderson, Will
Vaidyanathan, Ramanathan
Trau, Matt
Title Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing
Journal name Scientific Reports   Check publisher's open access policy
ISSN 2045-2322
Publication date 2015-01-06
Year available 2015
Sub-type Article (original research)
DOI 10.1038/srep07639
Open Access Status DOI
Volume 5
Start page 7639.1
End page 7639.7
Total pages 7
Place of publication London, United Kingdom
Publisher Nature Publishing Group
Language eng
Subject 1000 General
Abstract Exosomes are vesicles which have garnered interest due to their diagnostic and therapeutic potential. Isolation of pure yields of exosomes from complex biological fluids whilst preserving their physical characteristics is critical for downstream applications. In this study, we use 100 nm-liposomes from 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol as a model system as a model system to assess the effect of exosome isolation protocols on vesicle recovery and size distribution using a single-particle analysis method. We demonstrate that liposome size distribution and ζ-potential are comparable to extracted exosomes, making them an ideal model for comparison studies. Four different purification protocols were evaluated, with liposomes robustly isolated by three of them. Recovered yields varied and liposome size distribution was unaltered during processing, suggesting that these protocols do not induce particle aggregation. This leads us to conclude that the size distribution profile and characteristics of vesicles are stably maintained during processing and purification, suggesting that reports detailing how exosomes derived from tumour cells differ in size to those from normal cells are reporting a real phenomenon. However, we hypothesize that larger particles present in most purified exosome samples represent co-purified contaminating non-exosome debris. These isolation techniques are therefore likely nonspecific and may co-isolate non-exosome material of similar physical properties.
Formatted abstract
Exosomes are vesicles which have garnered interest due to their diagnostic and therapeutic potential. Isolation of pure yields of exosomes from complex biological fluids whilst preserving their physical characteristics is critical for downstream applications. In this study, we use 100 nm-liposomes from 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol as a model system as a model system to assess the effect of exosome isolation protocols on vesicle recovery and size distribution using a single-particle analysis method. We demonstrate that liposome size distribution and ζ-potential are comparable to extracted exosomes, making them an ideal model for comparison studies. Four different purification protocols were evaluated, with liposomes robustly isolated by three of them. Recovered yields varied and liposome size distribution was unaltered during processing, suggesting that these protocols do not induce particle aggregation. This leads us to conclude that the size distribution profile and characteristics of vesicles are stably maintained during processing and purification, suggesting that reports detailing how exosomes derived from tumour cells differ in size to those from normal cells are reporting a real phenomenon. However, we hypothesize that larger particles present in most purified exosome samples represent co-purified contaminating non-exosome debris. These isolation techniques are therefore likely nonspecific and may co-isolate non-exosome material of similar physical properties.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Art No. 7639

 
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Created: Fri, 16 Jan 2015, 20:43:03 EST by Mrs Louise Nimwegen on behalf of Aust Institute for Bioengineering & Nanotechnology