Open AccessImmunoaffinity‐based isolation of melanoma cell‐derived exosomes from plasma of patients with melanoma
Author(s) -
Sharma Priyanka,
Ludwig Sonja,
Muller Laurent,
Hong Chang Sook,
Kirkwood John M,
Ferrone Soldano,
Whiteside Theresa L.
Publication year - 2018
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1080/20013078.2018.1435138
Subject(s) - microvesicles , melanoma , exosome , flow cytometry , cancer research , liquid biopsy , cell , immunotherapy , medicine , immune system , cancer , immunology , chemistry , microrna , biochemistry , gene
Tumour‐derived exosomes (TEX) are a subset of extracellular vesicles (EVs) present in body fluids of patients with cancer. The role of this exosome subset in melanoma progression has been of interest ever since ex vivo studies of exosomes produced by melanoma cell lines were shown to suppress anti‐melanoma immune responses. To study the impact of melanoma‐derived exosomes (MTEX) present in patients’ plasma on melanoma progression, isolation of MTEX from total plasma exosomes is necessary. We have developed an immunoaffinity‐based method for MTEX capture from plasma of melanoma patients. Using mAb 763.74 specific for the CSPG4 epitope uniquely expressed on melanoma cells, we separated MTEX from non‐tumour cell‐derived exosomes and evaluated the protein cargo of both fractions by quantitative flow cytometry. Melanoma‐associated antigens were carried by MTEX but were not detectable in exosomes produced by normal cells. Separation of plasma‐derived MTEX from non‐MTEX provides an opportunity for future evaluation of MTEX as potential biomarkers of melanoma progression and as surrogates of melanoma in tumour liquid biopsy studies.