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. 2021 Nov 16;16(11):e0259563. doi: 10.1371/journal.pone.0259563

Fig 1. Isolation and separation of EVs by PVDF filtration and ultracentrifugation.

Fig 1

The culture medium of PANC-1 cells was pre-cleared by 10,000g centrifugation for 30 min at 4°C, and the resulting supernatant was passed through a 0.22μm PVDF centrifuge filter. The operational 3 (OP3) EVs (>220 nm) were trapped in the first filter. The filtered supernatant was then applied to a 0.1μm PVDF centrifuge filter. The operational 2 (OP2) EVs (100–220 nm) were trapped in the second filter and re-suspended in PBS from the filter surface. The operational 3 (OP1) EVs (<100 nm) in the final supernatant were recovered by ultracentrifugation (100,000g, 70 min at 4°C). For plasma EV isolation and separation, we started with the isolation of EVs using the Exoquick reagent. The resulting EVs were dissolved in PBS and applied to the filtration and separation process as depicted above.