Irvine CELLine Bioreactor Flask - page 3

CELLine Bioreactor Cell Culture
Figure 1A
Cell concentrations were measured every three to four days in the CELLine bioreactor
flasks. All four iterations were seeded with the same cell concentrations (1.5 x 10
6
cells/
mL). Cells were harvested, diluted 1:4 and reintroduced into the flask every 7 days.
Viable cell concentrations reached above 35 million cells/mL in each flask. This is
approximately 525 million cells in the CELLine 1000 flask and 175 Million cells in the
CELLine 350. At term, the CELLine bioreactors utilizing the BalanCD
CHO Growth A
medium maintained the highest cell concentrations.
Percent viability of cells in the CELLine bioreactor flasks were measured every three to four
days for 28 days. The viability slowly decreased over time; however, the concentration of
IgG was not affected. In the CELLine bioreactors, BalanCD
CHO Growth A maintained
the highest cell viability throughout the experiment.
Figure 1B
Shake Flasks Cell Culture
Shake flask cultures were monitored for viable cell density over the course of 14 days.
Viable cell density peaked with the fed-batch flask utilizing BalanCD
CHO Growth
A and the BalanCD
CHO Feed 1 above 10 million cells/mL. Both fed-batch cultures
maintained higher cell concentrations than the batch shake flask cultures.
Figure 1C
Percent viability of the cells in the shake flask cultures were monitored over the 14 day
experiment. Fed-batch cultures maintained higher percent viability than batch cultures
and were above 50% at term.
Figure 1D
Concentration g/L
Total Product
Harvest
Final
Yield
3
7
11
14
17
21
24
28
g/mL
Grams
C1000 Growth A
0.84
6.93
6.38
9.65
3.77
8.22
5.04
8.80
33.60 / 60
0.50
C350 Growth A
0.81
6.95
7.37
9.82
4.69
8.21
4.77
8.73
33.71 / 20
0.17
C1000 G10.7
0.96
6.68
6.85
9.62
4.17
7.61
5.17
7.54
31.45 / 60
0.47
C350 G10.7
0.92
6.50
6.92
9.53
4.48
7.54
4.52
6.89
30.46 / 20
0.15
Day
Condition
Antibody Production in CELLine Bioreactor Flask
Condition
Day
Total Product
Harvest
Total Yield
3
7
11
14
g/mL
Grams
Growth A Batch
0.15 1.18
1.52
1.63
1.63 / 30
0.049
Growth A / Feed 1 0.15 1.12
2.38
2.77
2.77 / 36
0.100
G10.7 Batch
0.16 1.05
1.29
1.36
1.36 / 30
0.041
G10.7 / Feed 3
0.16 1.29
2.67
2.78
2.78 / 36
0.100
Antibody Production in Shake Flask
Highlighted column indicates harvest
Highlighted column indicates harvest
The IgG concentration was measured throughout the CELLine bioreactor flask operation. A total of 60mL was collected from each CELLine 1000 over the course of 4 harvests from the
15mL cell compartment. A total of 20mL was collected from the CELLine 350 over the course of 4 harvests from the 5mL cell compartment. The results show up to 0.5 g IgG production
in 28 days and indicate linear scaling of the CELLine 1000 and CELLine 350 devices.
The shake flask IgG concentration was monitored during
culture for both batch and fed-batch conditions.
RESULTS
1,2 4,5,6
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