Extracellular vesicles separated from goat milk by differential centrifugation coupled with sodium citrate pretreatments

Satisfactory separation of milk-derived extracellular vesicles (MEVs) is important for the downstream analysis of the functions and properties of MEVs. However, the presence of abundant proteins in milk hindered the separation of MEVs. In this study, three pretreatment methods, including sodium citrate (SC), acetic acid (AA), and high-speed centrifugation, were adopted to separate MEVs from goat milk while minimizing the impact of protein. The MEVs were then characterized by nanoparticle tracking, transmission electron microscopy and western blotting experiments. The results indicated that pretreatments with AA and SC greatly decreased the impact of casein, but AA pretreatment damaged the surface structure of MEVs. Additionally, the differential centrifugation process resulted in a slight loss of MEVs. Overall, MEVs with small size and high purity can be obtained under 125 k × g centrifugation combined with SC pretreatment, which suggests a promising method for separation of MEVs from goat milk.