Isolation and analysis of extracellular vesicles from liquid biopsies
Research type
Research Study
Full title
Isolation and analysis of extracellular vesicles from liquid biopsies to gain valuable insights from patients and investigating their potential as biomarkers of disease
IRAS ID
348155
Contact name
Gayle Marshall
Contact email
Sponsor organisation
Medicines Discovery Catapult
Clinicaltrials.gov Identifier
N/A, N/A
Duration of Study in the UK
1 years, 9 months, 30 days
Research summary
Extracellular vesicles (EVs) are non-proliferative, lipid-bound vesicles that are secreted by most cell types. The function of EVs is not yet clear, however they are thought to play a role in cell-cell signaling and could be used as an important source of biomarkers. We aim to develop a workflow for isolating EVs from different sample types and assays to investigate the EV cargo from these samples for application to future projects, to better understand mechanisms underlying diseases and how EVs could be used as biomarkers.
Since EVs are secreted by most cell types, multiple sample types could be used to harvest and analyse these important biomarkers, including serum, plasma, CSF, urine and saliva samples. The proposed approach for this study is to investigate commercially available kits to isolate EVs from some of these samples, such as the Exo-spin mini columns. The isolated EVs will be analysed using dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). The cargo of the EVs will then be analysed using techniques such as Western blotting, Luminex assays and dSTORM to investigate the protein cargo within the EVs. Initial investigations into the quality of RNA cargo within the EVs will be conducted using the Agilent TapeStation and NanoDrop One platforms and if sufficient RNA quantity / quality is present, RNAseq could be used to analyse the RNA within the EVs. The effect of addition of the EVs isolated from these samples will be investigated in cell lines using commercially available kits such as the Incucyte Exofluor Green EV Labeling Kit, which can be used to label EVs and visualise the efficiency of their uptake by other cells and application of the previously described processes will be used to investigate the effect of EV uptake in the recipient cells.
REC name
HSC REC B
REC reference
24/NI/0131
Date of REC Opinion
15 Nov 2024
REC opinion
Further Information Favourable Opinion