Table 1.
Reusability of Nucleic Acids after Cargo Loading with pH-EVs
| Amount of siRNA in Loading Buffer (pmol) | Amount of pH-EVs Used (μg) | Amount of siRNA Loaded in EVs (pmol) | % of siRNA Loaded | Theoretical Unused siRNA (pmol) | siRNA Recovered after Washing (pmol) |
|---|---|---|---|---|---|
| 1,000 | 10 | 65 | 6.5 | 935 | 720 (filtrate 1) |
| 720 (filtrate 1) | 10 | 41 | 5.6 | 679 | 543 (filtrate 2) |
| 543 (filtrate 2) | 10 | 32 | 5.8 | 511 | 420 (filtrate 3) |
| 420 (filtrate 3) | 10 | 23 | 5.4 | 397 | 280 (filtrate 4) |
| 280 (filtrate 4) | 10 | 7 | 3.2 | 273 | 198 (filtrate 5) |
| 198 (filtrate 5) | 10 | ∼0 | ∼0 | ∼198 | 125 (filtrate 6) |
After incubation with pH-EVs, the amount of siRNA was calculated via fluorescence measurements. The remaining loading buffer (filtrate after washing loaded EVs) was then used to load a fresh set of EVs, and the measurements were repeated until RNA could no longer be loaded. Data were derived from three independent experiments with at least two technical replicates per experiment (n = 3).