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Comparative analysis of exosome isolation methods using culture supernatant for optimum yield, purity and downstream applications

Open access
Patel, Girijesh Kumar, Khan, Mohammad Aslam, Zubair, Haseeb, Srivastava, Sanjeev Kumar, Khushman, Moh’d, Singh, Seema, Singh, Ajay Pratap·2019·Scientific Reports

Abstract

Exosomes have received significant attention for their role in pathobiological processes and are being explored as a tool for disease diagnosis and management. Consequently, various isolation methods based on different principles have been developed for exosome isolation. Here we compared the efficacy of four kits from Invitrogen, 101Bio, Wako and iZON along with conventional ultracentrifugation-based method for exosome yield, purity and quality. Cell culture supernatant was used as an abundant source of exosomes, and exosome quantity, size-distribution, zeta-potential, marker-expression and RNA/protein quality were determined. The Invitrogen kit gave the highest yield but the preparation showed broader size-distribution likely due to microvesicle co-precipitation and had the least dispersion stability. Other preparations showed <150 nm size range and good stability. Preparation from iZON column; however, had a broader size-distribution in the lower size range suggestive of some impurities of non-vesicular aggregates. RNA quality from all preparations was comparable; however, proteins from Invitrogen method-based exosomal preparation showed polyethylene glycol (PEG) contamination in mass spectrometry. Chemical impurities from the precipitant could also be the cause of toxicity of Invitrogen method-based exosomal preparation in biological growth measurement assay. Together, these findings should serve as a guide to choose and further optimize exosome isolation methods for their desired downstream applications.

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Metadata

Authors
Patel, G. K., Khan, M. A., Zubair, H., Srivastava, S. K., Khushman, M., Singh, S., & Singh, A. P.
Venue
Scientific Reports
Published
2019-03-29
Volume
9
Issue
1
Pages
5335-5335
ISSN
2045-2322 (ISSN-L)
OpenAlex
W2925579059
Cited by
640
References
33
FWCI
22.49
Type
journal article
Language
en
Open access
gold

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CoChat. (2026). Citation check: Comparative analysis of exosome isolation methods using culture supernatant for optimum yield, purity and downstream applications. https://app.cochat.ai/papers/10.1038/s41598-019-41800-2
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Patel, G. K., Khan, M. A., Zubair, H., Srivastava, S. K., Khushman, M., Singh, S., & Singh, A. P. (2019). Comparative analysis of exosome isolation methods using culture supernatant for optimum yield, purity and downstream applications. Scientific Reports. https://doi.org/10.1038/s41598-019-41800-2

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