A Nanoparticle-Based Approach for the Detection of Extracellular Vesicles
Md. Khirul Islam; Urpo Lamminmäki; Saara Wittfooth; Kamlesh Gidwani; Kim Pettersson; Laura Lehtinen; Parvez Syed; Janne Leivo
A Nanoparticle-Based Approach for the Detection of Extracellular Vesicles
Md. Khirul Islam
Urpo Lamminmäki
Saara Wittfooth
Kamlesh Gidwani
Kim Pettersson
Laura Lehtinen
Parvez Syed
Janne Leivo
NATURE PUBLISHING GROUP
Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe2021042825090
https://urn.fi/URN:NBN:fi-fe2021042825090
Tiivistelmä
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.
Kokoelmat
- Rinnakkaistallenteet [19207]