Fig. 4. Flowchart of albumin depletion steps from human plasma-derived sEV-Ps, exomeres and supermeres.

Albumin spin columns are equilibrated in albumin binding buffer by centrifugation at 1,500g for 1 min at RT. The samples are then applied to the albumin-binding columns and incubated for 1 h at RT with rotation. The columns were centrifuged, and the albumin-depleted samples are collected in the flow- through. Albumin is eluted by centrifugation using albumin elution buffer. This protocol was modified from the Albumin Depletion Kit procedure (Abcam). The research conducted as part of this protocol complies with all the relevant ethical regulations. The use of the human samples was approved by the Vanderbilt University Medical Center Institutional Review Board (IRB; IRB nos 161529 and 151721).