Preprint Review Version 2 Preserved in Portico This version is not peer-reviewed

A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option?

Version 1 : Received: 20 July 2020 / Approved: 21 July 2020 / Online: 21 July 2020 (12:40:15 CEST)
Version 2 : Received: 19 August 2020 / Approved: 20 August 2020 / Online: 20 August 2020 (09:44:09 CEST)

A peer-reviewed article of this Preprint also exists.

Sidhom, K.; Obi, P.O.; Saleem, A. A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option? Int. J. Mol. Sci. 2020, 21, 6466. Sidhom, K.; Obi, P.O.; Saleem, A. A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option? Int. J. Mol. Sci. 2020, 21, 6466.

Journal reference: Int. J. Mol. Sci. 2020, 21, 6466
DOI: 10.3390/ijms21186466

Abstract

Extracellular vesicles (EVs) are membranous vesicles secreted by both prokaryotic and eukaryotic cells and play a vital role in intercellular communication. EVs are classified into several subtypes based on their origin, physical characteristics, and biomolecular makeup. Exosomes, a subtype of EVs, are released by the fusion of multivesicular bodies (MVB) with the plasma membrane of the cell. Several methods have been described in literature to isolate exosomes from biofluids including blood, urine, milk, and cell culture media among others. While differential ultracentrifugation (dUC), has been widely used to isolate exosomes, other techniques including ultrafiltration, precipitating agents such as poly-ethylene glycol (PEG), immunoaffinity capture, microfluidics and size exclusion chromatography (SEC) have emerged as credible alternatives with pros and cons associated with each. In this review, we provide a summary of commonly used exosomal isolation techniques with a focus on SEC as an ideal methodology. We evaluate the efficacy of SEC to isolate exosomes from an array of biological fluids, with a particular focus on its application to adipose tissue-derived exosomes. We argue that exosomes isolated via SEC are relatively pure and functional, and that this methodology is reproducible, scalable, inexpensive, and does not require specialized equipment or user expertise.

Subject Areas

extracellular vesicles; exosomes; microvesicles; differential ultracentrifugation; poly-ethylene glycol; immunoaffinity capture; microfluidics; size exclusion chromatography

Comments (1)

Comment 1
Received: 20 August 2020
Commenter: Ayesha Saleem
Commenter's Conflict of Interests: Author
Comment: We've made extensive edits as the manuscript undergoes revisions at IJMS, summarized below:
- completely updated and revised figure 1 and its legend
- added a new table 3
- section 2.6 on size exclusion chromatography has been modified - 75% new content
- section 3 revised - 30% new content
- added new section 4
- added new references
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