Barnes, Benjamin;
Caws, Thomas;
Thomas, Samantha;
Shephard, Alex P;
Corteling, Randolph;
Hole, Paul;
Bracewell, Daniel G;
(2022)
Investigating heparin affinity chromatography for extracellular vesicle purification and fractionation.
Journal of Chromatography A
, 1670
, Article 462987. 10.1016/j.chroma.2022.462987.
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Abstract
The purification of extracellular vesicles (EVs) remains a major hurdle in the progression of fundamental research and the commercial application of EV-based products. In this study, we evaluated the potential of heparin affinity chromatography (HAC) to purify neural stem cell-derived EVs as part of a multistep process. Bind-elute chromatography, such as HAC, is an attractive method of purification because it is highly scalable, robust and can be automated. Our findings support an interaction between EVs and heparin. The recovery of EVs using HAC based on particle counts was a minimum of 68.7%. We found HAC could remove on average 98.8% and 99.0% of residual protein and DNA respectively. In addition to EV purification, HAC was used to separate EVs into three populations based on their affinity to the heparin column. Within these populations, we detected differences in the expression of the exosome-associated protein TSG101 and the tetraspanin immunophenotype. However, the significance of these observations is not clear. Overall HAC shows promise as a potential purification method to capture EVs and this study proposes a novel application of HAC for EV fractionation. Moving forward, a better understanding of the heparin-EV interaction would be required before HAC can be more widely adopted for these applications.
| Type: | Article |
|---|---|
| Title: | Investigating heparin affinity chromatography for extracellular vesicle purification and fractionation |
| Location: | Netherlands |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.chroma.2022.462987 |
| Publisher version: | https://doi.org/10.1016/j.chroma.2022.462987 |
| Language: | English |
| Additional information: | Copyright © 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
| Keywords: | Exosomes, Extracellular vesicles, Heparin, Isolation, Purification |
| UCL classification: | UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering UCL > Provost and Vice Provost Offices > UCL BEAMS UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10146625 |
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