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

Edward GeeurickxJoeri TulkensBert DhondtJan Van DeunLien LippensGlenn VergauwenElisa HeyrmanDelphine De SutterKris GevaertFrancis ImpensIlkka MiinalainenPieter-Jan Van BockstalThomas De BeerMarca H M WaubenEsther N M Nolte-'t-HoenKatarzyna BlochJohannes V SwinnenEdwin van der PolRienk NieuwlandGeert BraemsNico CallewaertPieter MestdaghJo VandesompeleHannelore DenysSven EyckermanOlivier De WeverAn Hendrix
Published in: Nature communications (2019)
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.
Keyphrases
  • electronic health record
  • big data
  • machine learning
  • gene expression
  • genome wide
  • small molecule
  • dna methylation
  • molecularly imprinted
  • liquid chromatography
  • low cost
  • tandem mass spectrometry