Continuous Feeding Reduces the Generation of Metabolic Byproducts and Increases Antibodies Expression in Chinese Hamster Ovary-K1 Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Lines and Media
2.2. Process Conditions
2.3. Cells, Metabolites and Osmolality Analysis
2.4. Antibody Analysis by HPLC
2.5. Physicochemical Analysis
2.6. Statistical Analysis
3. Results and Discussion
3.1. Effects of Three Different Feeding Processes on CHO Cell Growth
3.2. Effects of Feeding Strategies on Cell Metabolism
3.3. Effects of Feeding Strategies on Antibody Expression and Quality
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Process | Feed Methods | Total CB7a Feeding Amount (V/V%) ① | Feeding Strategy | ||
---|---|---|---|---|---|
Cell Line A | Cell Line B | Cell Line A | Cell Line B | ||
Process 1 | Once every two days | 28 | 24 | 120 mL each time from Day 3 | |
Process 2 | Once every two days | 35 | 30 | 150 mL each time from Day 3 | |
Process 3 | Once every two days | 42 | 36 | 180 mL each time from Day 3 | |
Process 4 | Daily feeding | 28 | 24 | 60 mL each time from Day 3 | |
Process 5 | Daily feeding | 35 | 30 | 75 mL each time from Day 3 | |
Process 6 | Daily feeding | 42 | 36 | 90 mL each time from Day 3 | |
Process 7 | Continuous feeding ② | 28 | 24 | Starting from the third day, the on-time of the pump was set to 2S; the off-time was set to 738S | |
Process 8 | Continuous feeding | 35 | 30 | Starting from the third day, the on-time of the pump was set to 2S; the off-time was set to 590S | |
Process 9 | Continuous feeding | 42 | 36 | Starting from the third day, the on-time of the pump was set to 2S; the off-time was set to 491S |
Quality | Process 3 | Process 6 | Process 9 | |
---|---|---|---|---|
Cell line A | SEC purity (%) | 98.79 ± 0.43 | 99.05 ± 0.48 | 98.88 ± 0.54 |
CEX AV (%) | 24.60 ± 3.21 | 24.75 ± 2.88 | 22.70 ± 3.01 | |
CEX MP (%) | 66.08 ± 5.25 | 67.16 ± 4.72 | 68.23 ± 3.76 | |
CEX BV (%) | 9.32 ± 2.23 | 8.09 ± 1.85 | 9.06 ± 1.54 | |
G2F (%) | 0.27 ± 0.15 | 0.13 ± 0.13 | 0.33 ± 0.18 | |
G1F (%) | 7.80 ± 1.26 | 8.30 ± 1.83 | 7.53 ± 1.37 | |
G0F (%) | 86.63 ± 9.19 | 88.17 ± 8.38 | 89.37 ± 9.02 | |
High-mannose (%) | 2.15 ± 0.47 | 1.39 ± 0.43 | 0.88 ± 0.24 | |
NR-CE (%) | 96.12 ± 1.13 | 96.85 ± 1.56 | 96.52 ± 1.45 |
Quality | Process 3 | Process 6 | Process 9 | |
---|---|---|---|---|
Cell line B | SEC purity (%) | 97.75 ± 0.54 | 97.45 ± 0.50 | 98.05 ± 0.59 |
CEX AV (%) | 18.24 ± 2.34 | 19.23 ± 2.07 | 18.01 ± 1.34 | |
CEX MP (%) | 72.55 ± 4.54 | 72.01 ± 3.54 | 72.26 ± 3.29 | |
CEX BV (%) | 9.21 ± 1.47 | 8.76 ± 1.09 | 9.73 ± 1.21 | |
G2F (%) | 2.17 ± 1.38 | 1.89 ± 1.53 | 2.23 ± 1.77 | |
G1F (%) | 11.20 ± 1.88 | 13.15 ± 2.01 | 12.45 ± 2.34 | |
G0F (%) | 78.28 ± 8.78 | 77.58 ± 7.56 | 80.30 ± 9.57 | |
High-mannose (%) | 4.35 ± 1.43 | 3.38 ± 0.76 | 1.02 ± 0.35 | |
NR-CE (%) | 95.64 ± 1.02 | 95.43 ± 0.87 | 95.78 ± 0.93 |
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Xiao, S.; Ahmed, W.; Mohsin, A.; Guo, M. Continuous Feeding Reduces the Generation of Metabolic Byproducts and Increases Antibodies Expression in Chinese Hamster Ovary-K1 Cells. Life 2021, 11, 945. https://doi.org/10.3390/life11090945
Xiao S, Ahmed W, Mohsin A, Guo M. Continuous Feeding Reduces the Generation of Metabolic Byproducts and Increases Antibodies Expression in Chinese Hamster Ovary-K1 Cells. Life. 2021; 11(9):945. https://doi.org/10.3390/life11090945
Chicago/Turabian StyleXiao, Shang, Waqas Ahmed, Ali Mohsin, and Meijin Guo. 2021. "Continuous Feeding Reduces the Generation of Metabolic Byproducts and Increases Antibodies Expression in Chinese Hamster Ovary-K1 Cells" Life 11, no. 9: 945. https://doi.org/10.3390/life11090945