EV were isolated with a modified protocol of one described by Shin et al.20 (link). Isolation procedure was briefly abstracted in Fig. 6 .![]()
20 ml of plant lysate, cell culture media, blood serum, or parasite culture media were then centrifuged at 10,000 g for 10 minutes to remove larger contaminants. Supernatants were then passed through 0.22 μm filters to remove smaller contaminating particles, and to reduce the concentrations of larger extracellular vesicles. Filtered supernatants were then mixed at a 1:1 volume ratio with ATPS-EV isolation solution, and were then centrifuged at 1,000 g for 10 minutes for phase separation. Twice, 80% of the volume was removed from the upper, PEG-rich phase replaced with the upper phase of washing solution, prepared by mixing ATPS-EV isolation solution with distilled water at a 1:1 volume ratio and centrifuging at 1,000 g for 10 minutes, as described in Kim et al.21 (link).
Steps of the ATPS-based EV isolation protocol.
20 ml of plant lysate, cell culture media, blood serum, or parasite culture media were then centrifuged at 10,000 g for 10 minutes to remove larger contaminants. Supernatants were then passed through 0.22 μm filters to remove smaller contaminating particles, and to reduce the concentrations of larger extracellular vesicles. Filtered supernatants were then mixed at a 1:1 volume ratio with ATPS-EV isolation solution, and were then centrifuged at 1,000 g for 10 minutes for phase separation. Twice, 80% of the volume was removed from the upper, PEG-rich phase replaced with the upper phase of washing solution, prepared by mixing ATPS-EV isolation solution with distilled water at a 1:1 volume ratio and centrifuging at 1,000 g for 10 minutes, as described in Kim et al.21 (link).
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