Isolation methods commonly used to study the liposomal protein corona suffer from contamination issues

Kasper Kristensen, Rasmus Münter, Paul J Kempen, Mikkel E Thomsen, Allan Stensballe, Thomas L Andresen

Research output: Contribution to journalJournal articleResearchpeer-review

16 Citations (Scopus)

Abstract

The liposomal protein corona has been the focus of numerous studies, but there is still no consensus regarding its extent and composition. Rather, the literature is full of conflicting reports on the matter. To elucidate whether there could be a methodological explanation for this, we here scrutinize the efficiency of three commonly used liposome isolation methods at isolating stealth liposomes from human plasma. Firstly, we show that size-exclusion chromatography (SEC) in its standard form is prone to isolating unbound protein material together with the liposomes, but also that the method may be optimized to mitigate this issue. Secondly, we demonstrate that SEC in combination with membrane ultrafiltration is no better at removing the unbound protein material than SEC alone. Thirdly, we show that centrifugation is not able to pellet the liposomes. Overall, our results suggest that previous research on the liposomal protein corona may have suffered from significant methodological problems, in particular related to contaminant proteins interfering with the analysis of the protein corona. We believe that the data presented here may help guide future research around this challenge to reach a converging understanding about the properties of the protein corona on liposomes.

Original languageEnglish
JournalActa Biomaterialia
Volume130
Pages (from-to)460-472
Number of pages13
ISSN1742-7061
DOIs
Publication statusPublished - Aug 2021

Bibliographical note

Copyright © 2021 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Keywords

  • Isolation methods
  • Liposomes
  • Membrane ultrafiltration
  • Protein corona
  • Size-exclusion chromatography

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