Isolation of Bovine Milk Exosome Using Electrophoretic Oscillation Assisted Tangential Flow Filtration with Antifouling of Micro-ultrafiltration Membrane Filters

Abstract

Tangent flow-driven ultrafiltration (TF-UF) is an efficient isolation process of milk exosomes without morphological deformation. However, the TF-UF approach with micro-ultrafiltration SiN x membrane filters suffers from the clogging and fouling of microultrafiltration membrane filter pores with large bioparticles. Thus, it is limited in the long term, continuous isolation of large quantities of exosomes. In this work, we introduced electrophoretic oscillation (EPO) in the TF-UF approach to remove pore clogging and fouling of with micro-ultrafiltration SiN x membrane filters by large bioparticles. As a result, the combined EPO-assisted TF (EPOTF) filtration can isolate large quantities of bovine milk exosomes without deformation. Furthermore, several morphological and biological analyses confirmed that the EPOTF filtration approach could isolate the milk exosomes in high concentrations with high purity and intact morphology. In addition, the uptake test of fluorescent-labeled exosomes by the keratinocyte cells visualized the biological function of purified exosomes. Hence, compared to the TF-UF process, the EPOTF filtration produced a higher yield of bovine milk exosomes without stopping the filtering process for over 200 h. Therefore, this isolation process enables scalable and continuous production of morphologically intact exosomes from bovine milk, suggesting that high-quality exosome purification is possible for future applications such as drug nanocarriers, diagnosis, and treatments.

Extracellular Vesicles Comprising Carborane Prepared by a Host Exchanging Reaction as a Boron Carrier for Boron Neutron Capture Therapy

“…81,82 EVs were isolated from whey by ultracentrifugation 43,80,82,83 after EV concentration using tangential flow filtration, as reported previously. 84,85 Initially, the size distribution of EVs was evaluated by nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS) measurement. As a result, the mean and hydrodynamic diameters of EVs were 109 and 102 nm, respectively (Figure 2a, solid line; Table 1), and their size was favorable to achieving tumor accumulation via the EPR effect.…”

Recent research on material-based methods for isolation of extracellular vesicles

“…99 In addition, a study showed that combined electrophoretic oscillation-assisted tangent flow (EPOTF) filtration could isolate milk exosomes with high concentration, high purity and intact morphology. 100 Another method was designed by integrating a zwitterionic tandem membrane system, and one of the filters was modified with sulfobetaine methacrylate (SBMA) hydrogel to isolate exosomes. 101…”

Nature’s carriers: leveraging extracellular vesicles for targeted drug delivery

“…To date, there have been various separation, purification, and enrichment methods for collecting EVs, such as ultracentrifugation (Roerig et al., 2022 ), ultrafiltration (Ko et al., 2023 ), size exclusion chromatography (Lisi et al., 2023 ), polyethylene glycol precipitation (Jalaludin et al., 2023 ) and so on. Ultracentrifugation separates EVs based on the density, size, and shape of solutes.…”