FIGURE 7.

Impact of BPI (n = 3) on EV recovery, sample quality and ex vivo release of blood‐cell derived EV using different BCT (n = 5). Recovery analyses were performed on EV‐enriched fractions, separated from PDP by SEC, following 60 min (T1), 8 h (T2) and 72 h (T3) incubation of spiked rEV in whole blood at room temperature (n = 3). Sample quality and EV subtype analyses were performed on PDP, prepared 60 min (T1), 8 h (T2) and 72 h (T3) after blood draw (n = 5). Data are depicted as individual values with means. The asterisk indicates a statistically significant difference compared to the reference BPI (T1). a. Recovery rate (%) of recombinant EV (rEV), measured by fluorescent nanoparticle tracking analysis. b. Hemolysis, quantified as haemoglobin (Hb) concentration (mg/mL), measured by colorimetric assay. c. Platelet activation, quantified as plasma concentration of platelet factor 4 (PF4) (ng/mL), measured by ELISA. d. Platelet activation, quantified as plasma concentration of beta thromboglobulin (BTG) (ng/mL), measured by ELISA. e. Total EV count, quantified as the concentration (mL⁻¹) of particles with a refractive index < 1.42 (RI < 1.42). f. Activated platelet EV count, quantified as the concentration (mL⁻¹) of CD61+Lac+ particles (RI < 1.42). g. Non‐platelet EV count, quantified as the concentration (mL⁻¹) of CD61‐Lac+ particles (RI < 1.42). g. Residual platelet count, quantified as the concentration (mL⁻¹) of CD61+ particles (SSC cross section > 300 nm²). The dotted line indicates the detection limit of a standard haematology analyser. H. Erythrocyte EV count, quantified as the concentration (mL⁻¹) of CD235+ particles (RI < 1.42).