Figure 5.

A) Western Blot of RBC‐EVs, in contrast to plasma isolated EVs, show poor expression of tetraspanin triad CD63, CD9, and CD81, reflecting their biogenesis pathway (budding from RBC plasma membrane). Legend: M = Marker; Plasma EVs: EVs isolated from plasma by MSP‐agarose modified beads; RBC‐EVs = RBC EVs ectosomes. B) SiMoA beads were modified by MSP and by a combination of CD9/CD63/CD81 antibodies (Tetra) and used to capture serial dilutions of RBC‐EV using anti‐Band 3 antibody for detection. C) Detection signals reported as Average Enzyme per Bead (AEB) subtracted of the corresponding blank signals showing the capacity of MSP beads to capture RBC‐EVs down to 10⁸ vesicles/mL with a clear dose‐response trend which is not detectable neither by Tetra beads, nor by control (unfunctionalized) beads D) Detection of RBC‐EVs when spiked in plasma.