A1 Refereed original research article in a scientific journal
A Nanoparticle-Based Approach for the Detection of Extracellular Vesicles
Authors: Md. Khirul Islam, Parvez Syed, Laura Lehtinen, Janne Leivo, Kamlesh Gidwani, Saara Wittfooth, Kim Pettersson, Urpo Lamminmäki
Publisher: NATURE PUBLISHING GROUP
Publication year: 2019
Journal: Scientific Reports
Journal name in source: SCIENTIFIC REPORTS
Journal acronym: SCI REP-UK
Article number: ARTN 10038
Volume: 9
Number of pages: 9
ISSN: 2045-2322
DOI: https://doi.org/10.1038/s41598-019-46395-2
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/41363076
The analysis of extracellular vesicles (EVs) typically requires tedious and time-consuming isolation process from bio-fluids. We developed a nanoparticle-based time resolved fluorescence immunoassay (NP-TRFIA) that uses biotinylated antibodies against the proteins of tetraspanin family and tumorassociated antigens for capturing EVs from urine samples and cell culture supernatants without the need for isolation. The captured-EVs were detected either with Eu3+-chelate or Eu3+-doped nanoparticle-based labels conjugated either to antibodies against the tetraspanins or lectins targeting the glycan moieties on EVs surface. The NP-TRFIA demonstrated specific capturing and detection of EVs by antibodies and lectins. Lectin-nanoparticle based assays showed 2-10 fold higher signal-to-background ratio compared with lectin-chelate assays. The nanoparticle assay concept allowed surface glycosylation profiling of the urine derived-EVs with lectins. It was also applied to establish an assay showing differential expression of tumor-associated proteins on more aggressive (higher ITGA3 on DU145-and PC3-EVs) compared to less aggressive (higher EpCAM on LNCaP-EVs) PCa-cell lines derived-EVs. This NP-TRFIA can be used as a simple tool for analysis and characterization of EVs in urine and cell culture supernatants. Such approach could be useful in identification of disease-specific markers on the surface of patient-derived urinary EVs.
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