Characterization of end of production CD34+ cells transduced with TNS9.3.55 vector
| . | No. cells transduced . | Avg VCN liquid EC . | Frequency vector +ve CFUs (%) . | % CD34 d 3 . | Fold expansion d 0-d 3 . | No. CD34+ cells frozen . | CD34+ cells/kg . |
|---|---|---|---|---|---|---|---|
| Validation No. | |||||||
| 1 | 12.5 × 106 | 0.39 | 12.7 | 99.8 | 1.2 | 26 × 106 | NA |
| 2 | 300 × 106 | 0.63 | 30.0 | 98.0 | 1.5 | 500 × 106 | 10.3 × 106 |
| 3 | 54 × 106 | 0.58 | 23.8 | 98.7 | 1.7 | 106 × 106 | 2.1 × 106 |
| Mean ± SD | 0.53 ± 0.13 | 22.2 ± 8.8 | 98.8 ± 1.0 | 1.5 ± 0.25 | |||
| 2 (+23 mo) | NA | 0.56 | 12.7 | 99.0 | NA | NA | NA |
| . | No. cells transduced . | Avg VCN liquid EC . | Frequency vector +ve CFUs (%) . | % CD34 d 3 . | Fold expansion d 0-d 3 . | No. CD34+ cells frozen . | CD34+ cells/kg . |
|---|---|---|---|---|---|---|---|
| Validation No. | |||||||
| 1 | 12.5 × 106 | 0.39 | 12.7 | 99.8 | 1.2 | 26 × 106 | NA |
| 2 | 300 × 106 | 0.63 | 30.0 | 98.0 | 1.5 | 500 × 106 | 10.3 × 106 |
| 3 | 54 × 106 | 0.58 | 23.8 | 98.7 | 1.7 | 106 × 106 | 2.1 × 106 |
| Mean ± SD | 0.53 ± 0.13 | 22.2 ± 8.8 | 98.8 ± 1.0 | 1.5 ± 0.25 | |||
| 2 (+23 mo) | NA | 0.56 | 12.7 | 99.0 | NA | NA | NA |
Selected CD34+ HPC were either used fresh upon storage at 4°C overnight (validations 2 and 3) or thawed (validation 1). In validation 1, GLP vector stocks were used; in validations 2 and 3, vector stocks manufactured under cGMP were used. Two rounds of transduction at a multiplicity of infection of 25 were performed in presence of 100 ng/mL of TPO, Flt3-L, SCF and 20 U/mL interleukin-3. Approximately 80 CFUs per validation run were screened for the TNS9.3.55 by quantitative polymerase chain reaction. Avg, average. CFU, colony forming unit; EC, erythroid culture (d 14 bulk); NA, not applicable; SCF, stem cell factor; TPO, thrombopoietin.