Table 2.
Quantification of TNF, IL12 and PEG molecules loaded on gold nanoparticles
| Nanodruga | IL12 | TNF | PEG | |||
|---|---|---|---|---|---|---|
| IL12-ELISA | IL12-Bioassayd | TNF-Bioassaye | PEG-ELISA | |||
| IL12 (molecules/Np) | IL12 (molecules/Np) | EC50 (Nps/ml) | TNF (molecules/Np) | EC50 (Nps/ml) | PEG (molecules/Np) | |
| Bifunctional | ||||||
| iso1Au/IL12 | 6.4 ± 1.6 (n = 4)c | 47 ± 10 (n = 4) | 3.6 (± 0.8) × 106 (n = 4) | – | - | ~ 4 (n = 1) |
| iso1Au/TNF | – | – | – | 6.8 ± 1.0 (n = 3) | 52 (± 8) × 106 (n = 3) | ~ 3 (n = 1) |
| Trifunctional | ||||||
| iso1Au/TNF + IL12 (1:4.5) | 19.1 ± 3.8 (n = 5) | 35 ± 9 (n = 4) | 4.8 (± 1.2) × 106 (n = 4) | 2.4 ± 0.3 (n = 7) | 146 (± 22) × 106 (n = 7) | ~ 7 (n = 1) |
| iso1Au/TNF + IL12 (4:1) | 4.7 ± 0.6 (n = 8) | 40 ± 7 (n = 4) | 4.2 (± 0.7) × 106 (n = 4) | 14.1 ± 5.1 (n = 4) | 25 (± 9) × 106 (n = 4) | – |
| Proteinb | EC50 (molecules/ml) | EC50 (molecules/ml) | ||||
|---|---|---|---|---|---|---|
| IL12 | – | 169 (± 40) × 106 (n = 12) | – | – | – | |
| TNF | – | – | – | – | 352 (± 9) × 106 (n = 7) | – |
The italic numbers indicate the number of independent experiments
Mean ± SE
aPrepared as described in “Methods” section. The TNF:IL12 molar ratio used for iso1Au/TNF + IL12 preparation is indicated in parentheses
bRecombinant murine IL12 and TNF
cn, number of independent experiments
dIL12-induced secretion of IFNγ by murine splenocytes (reference standard, recombinant IL12) (see “Methods”). EC50: effective concentration 50
eL–M cell cytotoxicity assay (reference standard, recombinant TNF) [29]