Table 1.
Validation of LOD and confirmatory cut point.
| LOD determination—Mean and ratio | |||||
|---|---|---|---|---|---|
| Mean DMSO responses | Confirmatory response ratios | ||||
| Mean SFU/well | Response ratios: antigen vs DMSO | ||||
| Donor ID (n = 18) | Triplicate mean SFU/well | Triplicate mean SFU (log 10 transformed) | AAV5 pool 1/mock | AAV5 pool 2/mock | Transgene-product peptide pool/mock |
| 1 | 2 | 0.368 | 1.000 | 0.714 | 0.857 |
| 2C | 1 | 0.000 | 3.000 | 1.333 | 1.667 |
| 3D | 1 | 0.000 | 0.000 | 1.333 | 2.333 |
| 4 | 1 | 0.125 | 1.000 | 0.250 | 1.000 |
| 5 | 14 | 1.146 | 1.643 | 1.500 | 0.952 |
| 6 | 10 | 1.000 | 1.600 | 1.133 | 1.367 |
| 7A | 71 | 1.851 | 1.164 | 1.192 | 1.169 |
| 8 | 5 | 0.699 | 1.067 | 1.067 | 1.000 |
| 9 | 3 | 0.523 | 1.700 | 1.700 | 1.000 |
| 10B | 1 | 0.125 | 2.500 | 3.750 | 4.750 |
| 11 | 5 | 0.727 | 0.813 | 1.000 | 1.125 |
| 12 | 2 | 0.222 | 1.000 | 1.400 | 1.400 |
| 13 | 9 | 0.938 | 1.038 | 0.577 | 1.731 |
| 14 | 11 | 1.041 | 1.091 | 0.970 | 0.727 |
| 15 | 4 | 0.602 | 0.917 | 1.083 | 1.417 |
| 16 | 8 | 0.885 | 1.000 | 0.826 | 1.000 |
| 17 | 2 | 0.368 | 0.857 | 0.286 | 1.143 |
| 18 | 5 | 0.669 | 0.286 | 0.643 | 0.643 |
| Minimum | 0 | 0 | |||
| 25% percentile | 0.17 | 0.857 | |||
| Median | 0.60 | 1.000 | |||
| 75% percentile | 0.91 | 1.333 | |||
| Maximum | 1.15 | 1.731 | |||
| Mean | 0.56 | 1.049 | |||
| Std. deviation | 0.38 | 0.387 | |||
| LOD determination (normal distrib.) | Confirmatory cut point determination | ||||
| Mean + 1.645x StDev | 1.175 | Mean +1.645x StDev | |||
| LOD (Anti-log of above) | 15.0 | 1.69 | |||
Bold numbers represent the final calculated value for LOD and confirmatory cut point ratio respectively.
LOD and confirmatory cut point were based on a statistical analysis of the mean responses (SFU/well) under unstimulated (mock) conditions and the antigen/mock response ratios in naive monkeys, respectively. One result (7A (71)) was identified as a statistical outlier and excluded from the LOD analysis. Response ratios were obtained by dividing the triplicate mean SFU/well responses for AAV5 or transgene-product (TGP) peptide pools by the respective triplicate mean SFU/well response under unstimulated conditions (i.e., peptide stimulation/mock control). Five response ratios (2C (3.000), 3D (2.333) and 10B (2.500, 3.750 and 4.750)) were identified as statistical outliers and excluded from the confirmatory cut point analysis.