Comparison of separation methods for immunomodulatory extracellular vesicles from helminths

Abstract Helminths survive within their host by secreting immunomodulatory compounds, which hold therapeutic potential for inflammatory conditions. Helminth‐derived extracellular vesicles (EVs) are one such component proposed to possess activities. Due the recent discovery of helminth EVs, standardised protocols EV separation lacking. Excretory/secretory products porcine helminth, Ascaris suum , were used compare three methods: Size exclusion chromatography (SEC), ultracentrifugation (UC) and a combination two. Their performance was evaluated yield, sample purity ability EVs suppress lipopolysaccharide (LPS)‐induced inflammation in vitro. We found that all methods successfully separated with similar yield. Functional studies showed from reduced LPS‐induced levels tumour necrosis factor (TNF‐α) dose‐dependent manner. Overall, performance, however, UC+SEC presented slightly higher than either method alone.