For EVtrap characterization experiments, we used plasma from healthy individuals. For the kidney cancer analysis part of the project, we used 1 mL each of plasma from (a) five healthy individuals (no known kidney-related disease), (b) five patients with diagnosed CKD, some of whom were also diagnosed with kidney stones, and (c) five patients diagnosed with the most common form of kidney cancer, RCC. EVtrap beads were provided by Tymora Analytical as a suspension in water and were used as previously described in more detail.41 (link) The plasma samples were diluted 20 times in the diluent buffer, the EVtrap beads were added to the samples in a 1:2 v/ v ratio, and the samples incubated by end-over-end rotation for 30 min according to the manufacturer’s instructions. After supernatant removal using a magnetic separator rack, the beads were washed with PBS, and the EVs were eluted by a 10 min incubation with 200 mM triethylamine (TEA, Millipore-Sigma). The samples were fully dried in a vacuum centrifuge.