Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions

• Published in: Journal of extracellular vesicles Vol. 10; no. 10; pp. e12122 - n/a
• Main Authors: Vergauwen, Glenn, Tulkens, Joeri, Pinheiro, Cláudio, Avila Cobos, Francisco, Dedeyne, Sándor, De Scheerder, Marie‐Angélique, Vandekerckhove, Linos, Impens, Francis, Miinalainen, Ilkka, Braems, Geert, Gevaert, Kris, Mestdagh, Pieter, Vandesompele, Jo, Denys, Hannelore, De Wever, Olivier, Hendrix, An
• Format: Journal Article
• Language: English
• Published: United States John Wiley & Sons, Inc 01.08.2021; John Wiley and Sons Inc; Wiley
• Subjects: Biomarkers; Biomarkers - analysis; Blood; Breast cancer; Breast Neoplasms - blood; Breast Neoplasms - chemistry; Centrifugation; Centrifugation, Density Gradient - methods; Centrifuges; Chemical Fractionation - methods; Chromatography, Gel; corona; exosomes; Extracellular vesicles; Extracellular Vesicles - chemistry; Female; HIV Infections - blood; Humans; isolation; lipoprotein particles; Lipoproteins - analysis; miRNA; Ovarian cancer; Ovarian Neoplasms - blood; Ovarian Neoplasms - chemistry; Plasma; Plasma - chemistry; Protein composition; Proteome; proteomics; separation; transcriptomics; tRNA; California; United Kingdom > UK; Belgium; United States > US; Texas; Massachusetts; extracellular vesicles; proteomics; separation; biomarkers; exosomes; transcriptomics; isolation; lipoprotein particles; blood; corona
• ISSN: 2001-3078; 2001-3078
• DOI: 10.1002/jev2.12122

Separating extracellular vesicles (EV) from blood plasma is challenging and complicates their biological understanding and biomarker development. In this study, we fractionate blood plasma by combining size‐exclusion chromatography (SEC) and OptiPrep density gradient centrifugation to study clinical context‐dependent and time‐dependent variations in the biomolecular landscape of systemically circulating EV. Using pooled blood plasma samples from breast cancer patients, we first demonstrate the technical repeatability of blood plasma fractionation. Using serial blood plasma samples from HIV and ovarian cancer patients (n = 10) we next show that EV carry a clinical context‐dependent and/or time‐dependent protein and small RNA composition, including miRNA and tRNA. In addition, differential analysis of blood plasma fractions provides a catalogue of putative proteins not associated with systemically circulating EV. In conclusion, the implementation of blood plasma fractionation allows to advance the biological understanding and biomarker development of systemically circulating EV.