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Nat Commun. 2019 Jul 23;10(1):3288. doi: 10.1038/s41467-019-11182-0.

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

Author information

1
Laboratory of Experimental Cancer Research, Department of Human Structure and Repair, Ghent University, Ghent, 9000, Belgium.
2
Cancer Research Institute Ghent, Ghent, 9000, Belgium.
3
Department of Urology, Ghent University Hospital, Ghent, 9000, Belgium.
4
Department of Medical Oncology, Ghent University Hospital, Ghent, 9000, Belgium.
5
Department of Gynaecology, Ghent University Hospital, Ghent, 9000, Belgium.
6
Department of Biomolecular Medicine, Ghent University, Ghent, 9000, Belgium.
7
VIB Center for Medical Biotechnology, Ghent, 9000, Belgium.
8
VIB Proteomics Core, Ghent, 9000, Belgium.
9
Biocenter Oulu, University of Oulu, Oulu, 90220, Finland.
10
Laboratory of Pharmaceutical Process Analytical Technology, Department of Pharmaceutical Analysis, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, 9000, Belgium.
11
Department of Biochemistry and Cell Biology, Faculty of Veterinary medicine, Utrecht University, Utrecht, CM, 3584, The Netherlands.
12
Laboratory of Lipid Metabolism and Cancer, Department of Oncology, University of Leuven, Leuven, 3000, Belgium.
13
Laboratory of Experimental Clinical Chemistry, Amsterdam University Medical Center, Amsterdam University, Amsterdam, AZ, 1105, The Netherlands.
14
Biomedical Engineering & Physics, Amsterdam University Medical Center, Amsterdam University, Amsterdam, AZ, 1105, The Netherlands.
15
Vesicle Observation Center, Amsterdam University Medical Center, Amsterdam, AZ, 1105, The Netherlands.
16
Center of Medical Genetics, Ghent University, Ghent, 9000, Belgium.
17
Laboratory of Experimental Cancer Research, Department of Human Structure and Repair, Ghent University, Ghent, 9000, Belgium. An.Hendrix@ugent.be.
18
Cancer Research Institute Ghent, Ghent, 9000, Belgium. An.Hendrix@ugent.be.

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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