Table 1.
Summary of key parameters in human T cell biomanufacturing
| Biomanufacturing Parametersa,b,c | Scale (mL) | VCDmaxd (106 cells/mL) | TVC (106 cells) | Viabilityd (%) | μd(hr− 1) | Tdc(hr) | |
|---|---|---|---|---|---|---|---|
| Medium | AIM-V | 30 | 1.90 ± 0.11 | 56.9 ± 3.2 | 76.8 ± 2.5e | 0.030 ± 0.001 | 23.2 ± 0.8 |
| OpTmizer | 30 | 2.83 ± 0.12 | 84.8 ± 3.6 | 87.5 ± 1.4e | 0.030 ± 0.004 | 23.1 ± 2.9 | |
| ImmunoCult | 30 | 3.49 ± 0.52 | 104.6 ± 15.5 | 91.8 ± 1.1 | 0.041 ± 0.000 | 17.1 ± 0.2 | |
| Seed and stimulation | 1st stimulated Day 0 seed | 80 | 8.33 ± 0.11 | 666.0 ± 8.5 | 93.0 ± 1.4 | 0.053 ± 0.001 | 13.0 ± 0.2 |
| 1st stimulated Day 2 seed | 80 | 1.89 ± 0.04 | 151.2 ± 3.4 | 95.0 ± 1.4 | 0.029 ± 0.002 | 23.8 ± 1.5 | |
| 1st stimulated Day 5 seed | 80 | 0.55 ± 0.02 | 44.1 ± 2.0 | 96.0 ± 1.4 | 0.007 ± 0.001 | 99.5 ± 17.2 | |
| 2nd stimulated Day 0 seed | 80 | 3.98 ± 0.06 | 318.4 ± 4.5 | 93.0 ± 0.0 | 0.042 ± 0.002 | 16.7 ± 0.7 | |
| Scale-upf | T75 | 10 | 4.05 ± 0.13 | 40.5 ± 1.3 | 90.5 ± 0.7 | 0.034 ± 0.002 | 20.4 ± 0.9 |
| SF125 | 30 | 3.65 ± 0.18 | 109.4 ± 5.3 | 91.5 ± 2.1 | 0.041 ± 0.001 | 16.9 ± 0.4 | |
| Spinner | 80 | 3.98 ± 0.06 | 318.4 ± 4.5 | 93.0 ± 0.0 | 0.042 ± 0.002 | 16.7 ± 0.7 | |
| Bioreactor | 800 | 6.40 ± 0.46 | 5120.0 ± 367.7 | 94.0 ± 1.4 | 0.046 ± 0.002 | 15.2 ± 0.7 | |
Notes
aThe human T cell biomanufacturing was performed in spinner flasks for 4 days with basal medium of OpTmizer, feed of 30 U/mL IL-2 and 1st stimulation on Day 0 and Day 2, unless otherwise specified.
bSamples from multiple donors were used in the biomanufacturing process development, and T cells isolated from the same donor were used in the evaluation of one biomanufacturing parameter (i.e., medium, seed and stimulation, and scale-up). Significant difference between bioreactor runs in triplicate with p ≤ 0.05 was considered in the two-tailed t test. The results of cross donor comparison are presented in Fig. 3.
cAll data are presented as mean ± standard error of the mean (SEM).
dThe variation in cell growth parameters, such as VCD, viability and cell growth rate, was caused by the different biomanufacturing process parameters.
eThe viabilities of the T cells when reaching maximal VCD are presented. The relatively low harvest viability in the medium evaluation study was caused by an extended seed culture process in the early stage of process development, i.e. 6 days instead of 4 days.
fThe scale-up strategy was designed following the procedure described in Fig. 1.