Table 4.
Good manufacturing practice-compliant production process evaluation.
| Parameter | Limit | Con1 | Con2 | Con3 | Con4 |
|---|---|---|---|---|---|
| Viable cells/ml | ≥5.0 × 106 | 10.5 × 106 | 6.3 × 106 | 10.8 × 106 | 8.2 × 106 |
| Cell viability (%) | ≥75 | 96.7 | 92.8 | 96.8 | 96.9 |
| Sterility | No growth | No growth | No growth | No growth | No growth |
| Endotoxin (IU/ml) | ≤30 | ≤30 | ≤30 | ≤30 | ≤30 |
| Mycoplasma | Negative | Negative | Negative | Negative | Negative |
| CD4 (%) | ≥90.0 | 99.6 | 99.2 | 98.6 | 99.4 |
| CD25 (%) | ≥80.0 | 98.9 | 98.6 | 85.5 | 93.3 |
| CD127 (%) | ≥10.0 | 0.00 | 0.05 | 0.68 | 0.00 |
| CD8 (%) | ≤0.10 | 0.00 | 0.00 | 0.06 | 0.00 |
| CD19 (%) | ≤1.0 | 0.00 | 0.00 | 0.00 | 0.00 |
| CD56/16 (%) | ≤1.0 | 0.00 | 0.00 | 0.06 | 0.00 |
| Labeling check reagent PE (%) | ≤400 | ≤400 | ≤400 | ≤400 | ≤400 |
| Labeling check reagent APC (%) | ≤400 | ≤400 | ≤400 | ≤400 | ≤400 |
| Suppression at ratio 1 + 1 (%) | ≥80.0 | 99.5 | 81.4 | 98.2 | 99.9 |
| Suppression at ratio 1 + 5 (%) | ≥60.0 | 96.1 | 60.4 | 97.2 | 99.4 |
| Suppression at ratio 1 + 10 (%) | ≥50.0 | 85.4 | 52.5 | 61.6 | 83.2 |
Four successive regulatory T cell (Treg) productions were performed with healthy donor-derived leukapheresis products. Productions passed the indicated limits for release. Viable cells per milliliter indicate the total number of life retrieved Treg in the final Treg product. Viability indicates the percentage of life cells in the final Treg product. Sterility indicates bacterial growth contaminating the final Treg product. CD4, CD25, CD127, CD8, CD19, and CD56/CD16 indicate surface expression on the final Treg product. Labeling check reagent PE and APC indicate the remaining number of anti-CD3/anti-CD28 beads in the final Treg product. Suppression at ratio 1 + 1, 1 + 5, and 1 + 10 indicate the amount of suppression within the first generation of proliferating CD8+ responder cells by the final Treg product.