Performance of Blood Collection Tubes for Extracellular Vesicle Analysis

Extracellular vesicle (EV) analysis from blood samples is under intense investigation and may lead to the clinical implementation of EV analysis in health and disease. However, an urgent need exists to compare different types of blood collection tubes (BCT) and blood processing intervals (BPI) on EV performance metrics since these pre-analytical factors influence stability of circulating EV, and contamination by ex-vivo generated EV . This study systematically evaluated the impact of preserving (n=6) and non-preserving (n=5) BCTs and different BPIs (n=3). To enable robust and objective comparisons between BCTs and BPIs, performance metrics were developed reflecting EV recovery, sample quality, ex-vivo release of blood-cell derived EV subtypes and EV omics profile. These data allow an informed selection of the most optimal BCT and BPI for specific EV analysis based on the corresponding performance metrics profile. These results will support further methodological standardization of EV studies, and the proposed metrics can serve as a platform to guide future research on EV pre-analytics.