Table 1.
Time points and specifications for decisive in-process controls (IPC) and quality controls (QC).
| Parameter | IPC Time Point Acceptance Criteria |
QC Time Point Acceptance Criteria |
Method |
|---|---|---|---|
| CD3+ cell viability | Day 5 determined/declared |
Day 12 (at harvest) ≥80% viable cells among CD45+CD3+ cells |
Flow cytometry Ph. Eur. 2.7.24 |
| CD3+ cell percentage | Day 5 determined/declared |
Day 12 (at harvest) ≥80% CD3+ cells among viable CD45+ cells |
Flow cytometry Ph. Eur. 2.7.24 |
| Transduction Frequency | Day 5 ≥5% CAR+ cells among viable CD3+ cells |
Day 12 (at harvest) ≥10% CAR+ cells among viable CD3+ cells |
Flow cytometry Ph. Eur. 2.7.24 |
| Identity CD3+CAR+ cells/ Potency |
Day 5 CD20 CAR Detection Reagent bound to viable CD3+ cells |
Day 12 (at harvest) CD20 CAR Detection Reagent bound to viable CD3+ cells |
Flow cytometry Ph. Eur. 2.7.24 |
| Dose (CD3+CAR+ cells/mL) | Day 5 determined/declared |
Day 12 IMP according to cohort dosage deviation of 20% is acceptable |
Flow cytometry Ph. Eur. 2.7.24 |
| Appearance | / | Day 12 IMP slightly turbid infusion dispersion, primary package integrity |
Visual inspection |
| Endotoxin | Day 5 determined/declared |
Day 12 IMP <5 EU/mL |
LAL Method D Ph. Eur. 2.6.14 |
| Microbiological Examination | Day 10 negative-to-date |
day 12 IMP negative (result obtained post-release) |
Ph. Eur. 2.6.27 |
| Mycoplasma | Day 10 negative-to-date |
Day 12 IMP negative (result obtained post-release) |
Ph. Eur. 2.6.7 |
| Vector Copy Number | / | Day 12 (at harvest) <5 copies per transduced cell (result obtained post-release) |
Real-time qPCR |