Table 1. Titration of rAAV vector stocks and AAV RSMs using ITR2 or cassette (transgene or polyA) target sequences for qPCR.
| Sample ID | ITR2 qPCR (vg/ml) |
Expression cassette qPCR (transgene or pA) (vg/ml) |
Ratio ITR2/cassette | |
|---|---|---|---|---|
| Mean SD | Mean SD | |||
| AAVBAC028 | 5.60 ± 0.16 E+12 | 4.97 ± 3.3 E+12 |
1.13 | |
| AAVBAC016 | 6.88 ± 0.12 E+12 | 2.29 ± 0.78 E+12 |
3.00 | |
| Prod 12/2012 n°1 | 8.94 ± 2.06 E+12 | 6.77 ± 4.4 E+12 |
1.32 | |
| Prod 12/2012 n°2 | 9.45 ± 1.34 E+12 | 7.00 ± 4.9 E+12 |
1.35 | |
| AAVBAC017 | 1.03 ± 0.14 E+13 | 3.38 ± 0.92 E+12 |
3.06 | |
| AAVBAC010 | 1.14 ± 0.43 E+13 | 5.60 ± 3.08 E+12 |
2.04 | |
| AAVBAC040 | 1.15 ± 0.38 E+13 | 7.12 ± 2.18 E+12 |
1.62 | |
| BACTRANS002 | 1.33 ± 0.52 E+13 | 3.60 ± 1.33 E+12 |
3.69 | |
| AAVBAC009 | 1.67 ± 0.81 E+13 | 6.60 ± 2.42 E+12 |
2.53 | |
| BACTRANS003 | 2.66 ± 1.1 E+13 | 8.78 ± 0.63 E+12 |
3.03 | |
| Mean | 2.28 | |||
| AAV2RSM (lab1) | 1.68 ± 0.2 E+11 | 3.26 ± 0.2 E+10 |
5.10 | |
| AAV2RSM (lab2) | 1.50 ± 0.4 E+11 | 2.10 ± 0.6 E+10 |
7.10 | |
| Mean | 6.10 | |||
| AAV8RSM (lab1) | 1.93 ± 0.05 E+12 | 5.65 ± 0.09 E+11 |
3.40 | |
| AAV8RSM (lab2) | 1.30 ± 0.2 E+12 | 1.90 ± 0.5 E+11 |
6.80 | |
| Mean | 5.10 | |||
Titrations of RSMs were performed in two independent laboratoris (lab1, lab2). Mean ITR2/cassette ratios are indicated in bold letters for rAAV vector stocks, AAV2RSM, and AAV8RSM.
AAV, adeno-associated virus; ITR, inverted terminal repeats; RSM, reference standard materials; qPCR, quantitative polymerase chain reaction.