1) Zap-CHO media supplement improved the growth kinetics and productivity of CHO cells in six commercial media formulations. Cell growth increased on average 190% and maximum cell density increased 50% with Zap-CHO supplementation. 2) Productivity increased an average of 179% across the six media formulations when supplemented with Zap-CHO. Detailed analysis in one medium showed a 70% increase in productivity and 4 extra days of viable cell culture. 3) Zap-CHO is an effective animal-free supplement that optimizes CHO cell growth kinetics and productivity in commercial serum-free and chemically defined media.

1. Zap-CHO Media Supplement Optimizes the Growth
Kinetics and Productivity of CHO cells
Steve Pettit, Mary Ann Santos, Tanya Tanner InVitria, Cell Culture Development, info@invitria.com, 800-916-8311
Abstract Figure 1 Table1. Average increase in the performance of CHO cells grown in 6
commercial media supplemented with Zap-CHO.
Improving the growth and productivity of CHO cells is a major challenge in the Growth of CHO (5 days)
50
bioprocessing industry. We evaluated the benefit of Zap-CHO media supplement in No additive No Additive Zap-CHO(2x) 10% FBS
the culture of CHO cells. Zap-CHO is a defined animal-free protein supplement that (2 g/L)
40 10% FBS Growth 5 100 269 299
optimizes CHO culture in modern serum-free, animal-free, and CD media.
day
Viable cells/ml 105
Zap-CHO supplement was evaluated in six diverse commercial media. Zap-CHO 30
IVC 100 290 303
supplement improved the growth kinetics of CHO cells in every medium tested. Cell
growth, measured by Integral of Viable Cells (IVC), increased 98% to 372% across the six 20 Productivity 100 279 241
media and averaged 190%. Maximum peak cell density typically increased 50%. Zap-
CHO improved cell viability and extended the time of the batch by 2-4 days. Volumetric 10
productivity increased an average of 179% across the six (6) media formulations. Detailed Figure 4
kinetic analysis of CHO cells grown in a commercial medium with Zap-CHO showed an 0
increase in productivity of 70% and an extension of cell viability of 4 days.
A B C D
Commercial Medium
E F
Growth Kinetic of CHO Cells in a Medium
Figure 1. Log-phase growth of CHO K1 cells grown in 6 commercial media (A-F) supplemented with Zap-CHO. CHO cells were Supplemented with Zap-CHO
These data indicate that Zap-CHO is an effective media supplement that optimizes the seeded in shake-batch culture at 1.5x105 cells/ml. The viable cell count was determined on day5. Zap-CHO increased the growth 80
rate in every medium tested (Blue). Growth rate was similar to that achieved in the same medium with 10% FBS (Black). No additive
growth kinetics and productivity of CHO cells in modern media formulations. (0.5 g/L)
Introduction and experimental approach
Figure 2 60
Cells/ml 105
The benefit of Zap-CHO (InVitria) on log-phase cell growth, IVC (the Integrated Viable Relative Increase in IVC (day11)
400
40
Cell concentration), and volumetric productivity was examined in a series of experiments. No additive
Zap-CHO was evaluated in six diverse modern commercial media. The six media (2 g/L)
Relative IVC (No Additive=100)
included serum-free, protein-free and chemically defined (CD) formulations. In addition, 300 10% FBS
20
we performed detail growth analysis that confirmed the benefits of Zap-CHO on cell
growth, viability, and productivity.
200 0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Material and Methods 100
Days
Figure 4. Growth of CHO cells in Commercial Medium B supplemented with 0.5 g/L Zap-CHO. Cells grew to higher density and
Cells, media, and adaptation. Adherent dhfr- CHO line DP-12 clone 1934 produces a survived longer. Similar optimized growth kinetics was seen in the other commercial media supplemented with Zap-CHO.
humanized monoclonal antibody. This line was adapted to shake culture in 6 different
commercial serum- free formulations supplemented with 0.5% dFBS. 0
A B C D E F
Figure 5
Commercial Medium
Cells were maintained in media, 0.5% dFBS, MTX 5 µM in 125 ml shake flasks and washed Relative Increase in Producitivity with Zap-CHO
twice to remove FBS prior to seeding. Shake-batch cultures were performed in duplicate 4 ml Figure 2. Increase in IVC of CHO cells grown in 6 commercial media (A-F) supplemented with Zap-CHO. CHO cells were seeded
in shake-batch culture at 1.5x105 cells/ml. The relative IVC (Intergral of Viable Cell concentration) was determined on day 11.
200
cultures seeded at 1.5 x 105 viable cells/ml. Cells were seeded at 3.0 x 105 viable cells/ml Zap-CHO increased IVC of CHO cells in every medium tested (Blue). The increase in IVC often surpassed the same medium
supplemented with 10% FBS (Black). The value for medium E are: Zap-CHO=470, 10% FBS=750.
Relativive Productivity
in triplicate 125 ml shake flasks containing 30 ml of medium for detailed growth analysis. 150
Viable cell determination, IVCN, and volumetric productivity. The viable cell Figure 3
100
concentration was determined daily by a Guava PCA cell counter. IVC (the integral
Relative Increase in Productivity (day11)
viable cell count) was calculated from the sum of the daily viable cell counts. The 400
Relative Antibody Titer (No Additive=100)
concentration of antibody produced was determined by quantitative ELISA (Bethyl). No additive 50
300 (2 g/L)
10% FBS
250 0
No Additive (0.5x)
Conclusions 200
Figure 5. Increased productivity of CHO cells grown in commercial medium B supplemented with 0.5 g/L Zap-CHO. Values
are normalized so that the unsupplemented medium (yellow) is set to a value of 100. Volumetric productivity increased 70%.
150
Zap-CHO was evaluated for its ability to optimize the growth
kinetics and productivity of CHO cells. Zap-CHO substantially 100
improved growth rates, increased maximum cell density, extended 50
cell viability, and increased volumetric productivity. CHO growth 0
A B C D E F
and productivity improved in every medium tested. These results Commercial Medium
show that Zap-CHO is an effective media supplement that optimizes Figure 3. Increase in productivity of CHO cells grown in 6 commercial media (A-F) supplemented with Zap-CHO. CHO cells
www.InVitria.com
were seeded in shake-batch culture at 1.5x105 cells/ml. The relative volumetric productivity was determined on day1 Zap-CHO
1.
the growth kinetics and productivity of CHO cells. increased the volumetric productivity of CHO cells in every medium tested (Blue). The increase in productivity with Zap-CHO
surpassed that of 10% FBS (Black). The value for medium E are: Zap-CHO=825, 10% FBS=810.