Irvine CELLine Bioreactor Flask - page 4

Experimental Design
Experimental – CELLine
Media Tested
BalanCD
CHO Growth A
IS CHO-CD G10.7
Vessels Used
350 CELLine (350mL)
1000 CELLine (1000mL)
Seeding Density
1.5 x 10
6
cells/mL
Culture Protocols
28 days
Cell count & IgG quantification every 3-4 days
Complete media refreshed every 7 days, split cells 1:4
Control – Shake Flask
Media Tested
BalanCD
CHO Growth A (-/+) BalanCD
CHO Feed 1
IS CHO-CD G10.7 (-/+) BalanCD
CHO Feed 3
Vessel Used
125mL flask (30mL working vol.)
Seeding Density
0.3 x 10
6
cells/mL
Culture Protocols
14 days
Cell count daily and IgG quantification @ day 7, 10 and 14
Feed initial 5% working volume of feed to fed-batch conditions
on days 3-6
Seed
9 x 10
6
Cells
Day 3
Count
Sample IgG
Feed 5%
Day 4
Count
Feed 5%
Day 5
Count
Feed 5%
Day 6
Count
Feed 5%
Day 7
Count
Sample IgG
Day 10
Count
Sample IgG
Feed Glucose
Day 14
Count
Harvest IgG
Fed-Batch 125mL Shake Flask Process
CELLine 1000 Bioreactor Process
Seed
22.5 x 10
6
Cells
Day 3
Count
Sample IgG
Day 7
Count
Harvest IgG
Reduce Cells 1:4
Change Media
Day 10
Count
Sample IgG
Day 14
Count
Harvest IgG
Reduce Cells 1:4
Change Media
Day 17
Count
Sample IgG
Day 21
Count
Harvest IgG
Reduce Cells 1:4
Change Media
Day 24
Count
Sample IgG
Day 28
Count
Harvest IgG
Media Information
BalanCD
CHO Growth A is a chemically-defined growth medium, containing no animal-derived components. The formulation was
developed using Irvine Scientific’s Rational Culture Media Design
®
strategy with three model cell lines, and specifically designed to support
growth and productivity in a variety of CHO cells in both batch and fed-batch cultures.
IS CHO-CD G10.7 is part of Irvine Scientific’s CHO Media Development Kit. It is a chemically-defined growth media with no animal
derived components that was designed to help control metabolic overflow and support growth and productivity in a variety of CHO cells
in both batch and fed-batch cultures.
BalanCD
CHO Feed 1 and BalanCD
CHO Feed 3 are both chemically defined, animal component free and protein-free feed media
designed to provide cell culture nutrients for fed-batch applications in conjunction with BalanCD
CHO Growth A. These feeds have been
developed to promote cell growth and MAb production in a variety of CHO cells.
CELLine
Bioreactor Flask
Materials and Methods
Shake Flask Culture
Shake flasks (125mL) were used in conjunction with BalanCD
CHO Growth A or IS CHO-CD G10.7, 30mL initial working volume. Five percent feed of
BalanCD
CHO Feed 1 or BalanCD™ CHO Feed 3, was provided on days 3, 4, 5 and 6 for a total of 20% initial volume. Conditions: shake speed 120 RPM,
5% CO
2
, and 37°C humidified incubator. Cells were counted with Beckman Coulter Vi-Cell and IgG quantified by Forte’ Bio Octet kEQ. Initial seeding density
was 0.3 x 10
6
cells/mL.
CELLine Bioreactor Flask Culture
CELLine bioreactor flasks were used in conjunction with BalanCD
CHO Growth A or IS CHO-CD G10.7. Cells were split 1:4 every 7 days. Media was refreshed
every seven days. Conditions: 5% CO
2
, and 37°C humidified incubator. Cells were counted with Beckman Coulter Vi-Cell and IgG quantified by Forte’ Bio Octet
kEQ. Initial seeding density was 1.5 x 10
6
cells/mL.
1,2,3 5,6
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