Isolation
and purification of extracellular vesicles (EVs) from
plasma is essential to understand the EV circulation mechanism and
discover biomarkers for the early detection of diseases. However,
the size range of lipoprotein particles such as high density lipoprotein
(HDL), low density lipoprotein (LDL), and very low density lipoprotein
(VLDL) overlap that of EVs, making it difficult to remove lipoproteins
from EVs. Here, we propose a method for the high efficiency separation
of EVs in plasma using agarose gel electrophoresis based on their
differences in size and zeta potential properties. Electrophoresis
track assays revealed that EVs propagate more slowly than HDL but
more quickly than LDL and VLDL in 1% agarose gel with pH 7.4 Tris-Acetate-EDTA
(TAE) buffer. The size and morphology of the electrophoresis-recovered
products were characterized to be consistent with typical EVs. In
addition, the biological function of recovered EVs was investigated
with cell uptake tests. The feasibility of this method was further
verified with human plasma samples. In summary, this technique has
the potential to become a convenient and efficient approach for high-purity
EV separation.
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