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. 2024 May 31;11(29):2400533. doi: 10.1002/advs.202400533

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© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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A) SiMoA Beads were modified by MSP (left panel) and by a combination of antibodies directed against CD9/CD63/CD81tetraspanin (right panel). Single tetraspanin immune‐phenotyping of plasma EVs from 6 healthy donors was run in parallel on the two types of beads. B) For both settings and each sample, the total AEB (Average Enzyme per Bead) is calculated as the sum of single CD9/CD63/CD81 AEB detection. Single tetraspanin expression level is calculated as the AEB% over the total AEB. Both methods confirm the expected heterogeneity of EV samples with remarkable accordance of the two systems in terms of differential tetraspanin profiling. AEB data and %CV are reported in Table S1 (Supporting Information).