University of Oulu

Geeurickx, E., Tulkens, J., Dhondt, B. et al. The generation and use of recombinant extracellular vesicles as biological reference material. Nat Commun 10, 3288 (2019). https://doi.org/10.1038/s41467-019-11182-0

The generation and use of recombinant extracellular vesicles as biological reference material

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Author: Geeurickx, Edward1,2; Tulkens, Joeri1,2; Dhondt, Bert1,2,3;
Organizations: 1Laboratory of Experimental Cancer Research, Department of Human Structure and Repair, Ghent University, Ghent, 9000, Belgium
2Cancer Research Institute Ghent, Ghent, 9000, Belgium
3Department of Urology, Ghent University Hospital, Ghent, 9000, Belgium
4Department of Medical Oncology, Ghent University Hospital, Ghent, 9000, Belgium
5Department of Gynaecology, Ghent University Hospital, Ghent, 9000, Belgium
6Department of Biomolecular Medicine, Ghent University, Ghent, 9000, Belgium
7VIB Center for Medical Biotechnology, Ghent, 9000, Belgium
8VIB Proteomics Core, Ghent, 9000, Belgium
9Biocenter Oulu, University of Oulu, Oulu, 90220, Finland
10Laboratory of Pharmaceutical Process Analytical Technology, Department of Pharmaceutical Analysis, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, 9000, Belgium
11Department of Biochemistry and Cell Biology, Faculty of Veterinary medicine, Utrecht University, Utrecht, CM, 3584, The Netherlands
12Laboratory of Lipid Metabolism and Cancer, Department of Oncology, University of Leuven, Leuven, 3000, Belgium
13Laboratory of Experimental Clinical Chemistry, Amsterdam University Medical Center, Amsterdam University, Amsterdam, AZ, 1105, The Netherlands
14Biomedical Engineering & Physics, Amsterdam University Medical Center, Amsterdam University, Amsterdam, AZ, 1105, The Netherlands
15Vesicle Observation Center, Amsterdam University Medical Center, Amsterdam, AZ, 1105, The Netherlands
16Center of Medical Genetics, Ghent University, Ghent, 9000, Belgium
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 2.8 MB)
Persistent link: http://urn.fi/urn:nbn:fi-fe2020042019296
Language: English
Published: Springer Nature, 2019
Publish Date: 2020-04-20
Description:

Abstract

Recent years have seen an increase of extracellular vesicle (EV) research geared towards biological understanding, diagnostics and therapy. However, EV data interpretation remains challenging owing to complexity of biofluids and technical variation introduced during sample preparation and analysis. To understand and mitigate these limitations, we generated trackable recombinant EV (rEV) as a biological reference material. Employing complementary characterization methods, we demonstrate that rEV are stable and bear physical and biochemical traits characteristic of sample EV. Furthermore, rEV can be quantified using fluorescence-, RNA- and protein-based technologies available in routine laboratories. Spiking rEV in biofluids allows recovery efficiencies of commonly implemented EV separation methods to be identified, intra-method and inter-user variability induced by sample handling to be defined, and to normalize and improve sensitivity of EV enumerations. We anticipate that rEV will aid EV-based sample preparation and analysis, data normalization, method development and instrument calibration in various research and biomedical applications.

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Series: Nature communications
ISSN: 2041-1723
ISSN-E: 2041-1723
ISSN-L: 2041-1723
Volume: 10
Article number: 3288
DOI: 10.1038/s41467-019-11182-0
OADOI: https://oadoi.org/10.1038/s41467-019-11182-0
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Subjects:
Funding: This work was supported by Ghent University (Concerted Research Actions and Industrial Research Fund), Ghent University Hospital, Kom Op Tegen Kanker, postdoctoral position and Krediet aan Navorsers (A.H.) and PhD position strategic basic research (E.G.) from Fund for Scientific Research Flanders (FWO), by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 722148 and by the European Research Council under the European Union’s Seventh Framework Programme [FP/2007–2013]/ERC Grant Agreement number [337581]. E.V.P.D. was funded by the Netherlands Organisation for Scientific Research—Domain Applied and Engineering Sciences (NWO-TTW), research program VENI 15924.
Copyright information: © The Author(s) 2019. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
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