Table 2. Inputs and Outputs from Iteration B.
| experimental
inputs | |||
|---|---|---|---|
| factor | levels | ||
| phospholipid content (mol %) | 12.5 | 15 | 17.5 |
| ionizable lipid type (corresponding pKa) | D-Lin-MC3-DMA (6.4) | ALC-0315 (6.09) | SM-102 (6.75) |
| ionizable lipid content (mol %) | 35 | 40 | 45 |
| aqueous-phase pH | 4 | 5 | 6 |
| Fixed Parameters | |||
| N/P ratio | 10 | ||
| phospholipid type | DOPE | ||
| DMG-PEG-2000 content (mol %) | 1.25 | ||
| total flow rate (mL/min) | 16 | ||
| ambient temperature during formulation (°C) | 20 | ||
| experimental
outputs | |
|---|---|
| critical quality attributes | analytical method |
| size | dynamic light scattering |
| PDI | dynamic light scattering |
| EE | RiboGreen Assay |
| charge | dynamic light scattering |
| % filled particles | RiboGreen assay/nanoparticle tracking analysis |
| % full RNA transcripts | BioAnalyzer |
| protein expression | luciferase assay |
| cellular activation | MSD cytokine assay |