Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids

Chandrasekera, Dhananjie; Shah, Rishi; van Hout, Isabelle; De Jonge, Willow; Bunton, Richard; Parry, Dominic; Davis, Philip; Katare, Rajesh

doi:10.7150/ntno.82939

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Nanotheranostics 2023; 7(4):345-352. doi:10.7150/ntno.82939 This issue Cite

Research Paper

Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids

Dhananjie Chandrasekera¹, Rishi Shah¹, Isabelle van Hout¹, Willow De Jonge¹, Richard Bunton², Dominic Parry², Philip Davis², Rajesh Katare^1✉

1. Department of Physiology, HeartOtago, School of Biomedical Sciences, University of Otago, Dunedin, New Zealand.
2. Cardiothoracic Surgery, Dunedin School of Medicine, University of Otago, Dunedin, New Zealand.

Citation:

Chandrasekera D, Shah R, van Hout I, De Jonge W, Bunton R, Parry D, Davis P, Katare R. Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids. Nanotheranostics 2023; 7(4):345-352. doi:10.7150/ntno.82939. https://www.ntno.org/v07p0345.htm

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Abstract

Extracellular vesicles (EVs), such as exosomes, are nanovesicles that have received significant attention due to their ability to contain various molecular cargos. EVs found in biological fluids have been demonstrated to have therapeutic potential, including as biomarkers. Despite being extensively studied, a significant downfall in EV research is the lack of standardised protocol for its isolation from human biological fluids, where EVs usually exist at low densities. In this study, we tested two well-established EV isolation protocols, precipitation, and size exclusion chromatography (SEC), to determine their efficiency in isolating EVs from the pericardial fluid. Precipitation alone resulted in high yields of low-purity exosomes as tested by DLS analysis, transmission electron microscopy, immunogold labelling and western blotting for the exosomal surface proteins. While EVs isolated by SEC were pure, the concentration was low. Interestingly, the combination of precipitation followed by SEC resulted in high EV yields with good purity. Our results suggest that the combination method can be adapted to isolate EVs from body fluids which have low densities of EV.

Keywords: Exosomes, Extracellular vesicles, Biofluids, Precipitation, Size-exclusion chromatography

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APA

Chandrasekera, D., Shah, R., van Hout, I., De Jonge, W., Bunton, R., Parry, D., Davis, P., Katare, R. (2023). Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids. Nanotheranostics, 7(4), 345-352. https://doi.org/10.7150/ntno.82939.

ACS

Chandrasekera, D.; Shah, R.; van Hout, I.; De Jonge, W.; Bunton, R.; Parry, D.; Davis, P.; Katare, R. Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids. Nanotheranostics 2023, 7 (4), 345-352. DOI: 10.7150/ntno.82939.

NLM

CSE

Chandrasekera D, Shah R, van Hout I, De Jonge W, Bunton R, Parry D, Davis P, Katare R. 2023. Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids. Nanotheranostics. 7(4):345-352.

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