Figure 5
Dynamic range and limit of detection. EVs were captured on anti-CD63-coated beads followed by detection with biotinylated anti-CD9 antibody (Clone VJ1/20). (A) Flow cytometry analysis profiles. Increasing amounts (30 ng to 10 μg; 1.37∙107–4.59∙109 particles) of PC3-derived EVs were captured on 6 μm beads and detected by flow cytometry. The total volume of the assay was 100 μl. The graph represents the overlay of the curves obtained. The legend indicates the amount of EVs and the RFI for each curve. (B) Regression analysis. RFI (right) and SI values (left) were plotted as a scatter graph and fitted to a polynomic curve (upper panel). Linearity can be observed between 0.2–8 µg of PC3-derived EVs with a r2 > 0.98 (lower panels). The minimal amount of EVs detected (RFI > 1), corresponds to 30 ng yielding a RFI of 1.08. A representative experiment out of 3 is shown.
