Download Report

Cell Culture
Get the most from your
fed-batch cultures
GIBCO CHO CD EfficientFeed™ Kit
®
Cell Culture
Get the most from your fed-batch cultures
GIBCO CHO CD EfficientFeed™ Kit
®
→→ Demonstrated 2- to 5-fold increases in productivity
→→ Identify optimal feed formulation to grow cells faster for bioreactor seeding
→→ Chemically defined—no growth factors, lysates, or less consistent hydrolysates needed
→→ Made from AGT™ powders plus supplements, for simplified transition to larger scales
The CHO CD EfficientFeed™ Kit is a system designed to help you
The CHO CD EfficientFeed™ Kit includes:
more quickly and efficiently identify a feed supplement formulation
→→ One liter of CHO CD EfficientFeed™ A—a chemically defined
that will improve fed-batch productivity 2- to 5-fold compared to
formulation reconstituted from AGT™ powders plus supple-
batch conditions, as well as help you identify a formulation that will
ments. It contains a carbon source, concentrated amino acids,
help grow bioreactor cell seed stocks more quickly.
vitamins, and trace elements that can boost cell growth and
productivity alone or in combination with CHO CD Efficient-
Efficiency is getting two benefits from one kit
Feed™ B. This product contains no lipids, hydrolysates, or
The CHO CD EfficientFeed™ Kit contains two feed supplements, A
growth factors to achieve its results.
and B, based on successful chemically defined media you may be
→→ One liter of CHO CD EfficientFeed™ B—a second chemically
using or considering for your batch culture medium (CD OptiCHO™
defined formulation reconstituted from AGT™ powders plus
Medium or CD CHO Medium). Prepared using chemically defined
supplements. It also contains a carbon source, concentrated
components in higher concentrations, and based on typical CHO
amino acids, vitamins, and trace elements that can boost
culture needs, these feeds demonstrate the capacity for increas-
cell growth and productivity alone or in combination with
ing production of proteins and antibodies 2- to 5-fold compared
CHO CD EfficientFeed™ A. This product contains no lipids,
to batch culture conditions (Figures 1 and 2). Gain increased pro-
hydrolysates, or growth factors to achieve its results.
ductivity from one formulation and higher cell growth kinetics
→→ A manual that guides you through decisions related to
from the other, or a combination of the two. Higher cell growth
chemical and physical compatibility as well as options for
kinetics provide quicker growth of cells for seeding larger-scale
shake flask or benchtop bioreactor experimentation to help
bioreactors. Being able to load the bioreactor more quickly may
you select the best feeds for your situation and particular
allow you to increase the number of bioreactor runs possible
CHO cell clone.
within a year (Figure 3). The kit provides a complete workflow and
the choice to either use recommended protocols or develop your
own protocols for cell growth or protein production.
2
CD OptiCHO™ batch
Fed with 15% A on day 0 and 10% A on day 3
Fed with 30% A on day 0
Fed with 30% A/B mixture on day 0
CD OptiCHO™ batch, IgG
Fed with 15% A on day 0 and 10% A on day 3, IgG
Fed with 30% A on day 0, IgG
Fed with 30% A/B mixture on day 0, IgG
CD OptiCHO™ batch
Fed with 15% A on day 0 and 10% A on day 3
Fed with 30% A on day 0
Fed with 30% of A/B mixture on day 0
CD OptiCHO™ batch, IgG
Fed with 15% A on ay 0 and 10% A on day 3, IgG
Fed with 30% A on day 0, IgG, IgG
Fed with 30% of A/B mixture on day 0
5,000
3,000
2,000
5-fold
increase
106
1,000
2-fold
increase
0
2
4
6
8
10
12
14
16
107
4,000
3,000
6-fold increase
2,000
106
2.5-fold increase
0
0
Time (days)
Figure 1—Fed-batch culture using CD OptiCHO™ as base medium. Cells were grown
in CD OptiCHO™ Medium for three passages prior to this study. Cultures were
seeded in triplicate at 3 × 105 viable cells/ml in 50 ml in 250 ml shake flasks.
Cultures were incubated at 37°C, 8% CO2, and shaken at 125 rpm. Different
fed-batch strategies are described in the legend. No further supplementation
was performed during culture. Viable cell densities were determined using a
Coulter Vi-CELL™ analyzer. IgG levels were determined by protein affinity HPLC.
The IgG CHO cell line was developed by Chromos using ACE technology.
IgG (mg/L)
4,000
Viable cell density (cells/ml)
107
IgG (mg/L)
Viable cell density (cells/ml)
5,000
2
4
6
8
10
12
14
16
1,000
0
Time (days)
Figure 2—Fed-batch culture using CD OptiCHO™ as base medium (recombinant IgG
clone #2). Cells were grown in CD OptiCHO™ Medium for three passages prior
to the study. Cultures were seeded in triplicate at 3 × 105 viable cells/ml in
50 ml in 250 ml shake flasks. Cultures were incubated at 37°C, 8% CO2, and
shaken at 125 rpm. Different fed-batch strategies are described in the legend. No further supplementation was performed during culture. Viable cell
densities were determined using a Coulter Vi-CELL™ analyzer. IgG levels were
determined by protein affinity HPLC. The IgG CHO cell line was developed by
Chromos using ACE technology.
Without cell-mass feeding strategy:
7 days × 3 scale-ups (on average) before bioreactor = 21 days to prepare for run
With cell-mass feeding strategy:
6 days × 3 scale-ups (on average) before bioreactor = 18 days to prepare for run
If 12 bioreactor runs (on average) per year:
• Could save 36 days of prep time
• Might allow at least 1 more run per year
Figure 3—Cell growth feeding benefits. Results will vary based on specific cell clone and bioreactor type.
3
www.invitrogen.com
Faster scale-up capability
Simplify your path from small-scale experiments to larger-scale
production runs using CHO CD EfficientFeed™ Supplements made
from AGT™ powders plus supplements. Furthermore, experiments
have shown scalability from shake flasks to bioreactors (Figure 4).
So you can have confidence that when you order larger quantities
in more economical formats, like AGT™, your results will be similar
DASGIP bioreactors
Shake flasks
to those with liquids tested from this kit.
1,800
Customize your results
to meet your specific needs
1,200
900
Whether you choose to do some simple testing of each CHO CD
600
300
EfficientFeed™ supplement alone, in combination with each other,
0
or in combination with a growth factor or hydrolysate, this kit and
Ba
tc
1
h
on 0%
da C
10
ys HO
%
3, F
1:
6, eed
1
an A
C
d
on HO
8
da Fe
ys ed
10
3, A a
%
6, nd
1:
an B
1
d
on CH
8
da O
F
ys e
e
2, d
A
4,
6, and
an B
d
8
Fla
sk
ba
tc
h
1
on 0%
da C
ys HO
10
%
3, F
6, eed
1:
1
an A
CH
d
on O
8
da Fe
ys ed
10
3, A a
%
6, nd
1:
an B
1
d
on CH
8
da O F
ys ee
2, d A
4,
6, and
an B
d
8
IgG (mg/L)
1,500
Figure 4—Representative day 11 IgG levels in bioreactors and shake flask controls.
Chromos CHO clone #1, cells that have been growing in CD OptiCHO™ Medium
for over three passages, were inoculated at 3 × 105 viable cells/ml in DASGIP (0.5 L
working volume) and in 250 ml shake flasks (60 ml working volume). Bioreactors
were controlled at 37°C, pH 7.05, with direct oxygen at 50%. Shake flask cultures
were conducted as previously described. Different feed combinations and feeding strategies were tested. IgG levels were determined by protein affinity HPLC.
manual contain the information and guidance to help you succeed
in boosting your cell growth and productivity levels. They also will
help you identify when additional support from our PD-Direct™
Services team would be useful. When you want basal formulation
customization, combined feed formulation customization, custom
packaging, or production in AGT™ format, contact the GIBCO®
Custom Product Services team with your particular requirements.
Ordering information
Product Quantity Cat. no.
1 kit A10241-01
CHO CD EfficientFeed™ A
1,000 ml
A10234-01
CHO CD EfficientFeed™ B
1,000 ml
A10240-01
CHO CD EfficientFeed™ Kit
Kit components, also available separately:
Results here are based on use of the following basal media. Consider them to start your process and feed optimization efforts:
CD OptiCHO™ Medium 1,000 ml 12681-011
CD CHO Medium
1,000 ml
10743-029
For more information, call your regional Customer Service representative or visit www.invitrogen.com/bioproduction.
www.invitrogen.com
©2007 Invitrogen Corporation. All rights reserved. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the
terms and conditions of all applicable Limited Use Label Licenses. For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. B-073176-r1 1007