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. 2021 Jun 9;22(12):6234. doi: 10.3390/ijms22126234

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Immune capture with biotinylated anti-CSPG4 Abs of melanoma cell-derived exosomes on streptavidin-labeled beads from plasma of patients with melanoma. Exosomes were first isolated from plasma by size exclusion chromatography (SEC; step #1). Fraction #4 containing a mix of melanoma cell-derived vesicles (MTEX) and vesicles derived from non-malignant cells (NMTEX) were immuno-captured (step #2) Non-captured exosomes (NMTEX) in red were re-captured on beads with anti-CD63 mAbs for detection by on-bead flow cytometry (step #3). Two different clones of CSPG4 mAb (763.74 and 225.28) specific for distinct epitopes of this protein were used for capture and detection, respectively. Reproduced with changes from Sharma et al., Ref # 58.