Table 1.
Single and combination adjuvant loading on PLGA NP and doses for GM-CSF and FLT3L BMDCs in isoMLR assays. Size, PDI and zeta measurements were taken for all NPs prior to electrostatically loading adjuvants CpG or PUUC. PDI is polydispersity index. *The target adjuvant concentration for R848 was 6 μg per mg PLGA. Due to batch-to-batch variation, encapsulation of R848 in PLGA NP ranged between 4.8 and 6.67 μg per mg NP and the adjuvant dose ranged between 80 to ~110 ng per 500,000 BMDCs.
| PLGA-PEI NP Formulation | Size (nm) | PDI | Zeta (mV) | Targeted adjuvant concentration |
Adjuvant dose for BMDCs |
|---|---|---|---|---|---|
| (per mg NP) | (ng/5e5 cells/mL) | ||||
| CpG NP | 269.4 ± 41.1 | 0.45 | 31.7 ± 0.7 | 60 μg CpG | 500 ng CpG |
| MPLA NP | 246.0 ± 69.26 | 0.134 | 25.0 ± 4.92 | 6 μg MPLA | 100 ng MPLA |
| PUUC NP | 269.4 ± 41.1 | 0.45 | 31.7 ± 0.7 | 6 μg PUUC | 100 ng PUUC |
| R848 NP | 253.2 ± 61.10 | 0.37 | 29.1 ± 2.3 | 4.8 μg R848 | 100 ng R848 |
| CpG+PUUC NP | 269.4 ± 41.1 | 0.45 | 31.7 ± 0.7 | 30 μg CpG and 6 μg PUUC |
500 ng CpG and 100 ng PUUC |
| MPLA+CpG NP | 246.0 ± 69.26 | 0.134 | 25.0 ± 4.92 | 6 μg MPLA and 60 μg CpG |
50 ng MPLA and 500 ng CpG |
| MPLA+PUUC NP | 246.0 ± 69.26 | 0.134 | 25.0 ± 4.92 | 6 μg MPLA and 6 μg PUUC |
100 ng MPLA and 100 ng PUUC |
| R848+PUUC NP | 253.2 ± 61.10 | 0.37 | 29.1 ± 2.3 | 6 μg R848* and 6 μg PUUC |
100 ng R848* and 100 ng PUUC |