Abstract
In laboratory scale therapeutical protein production, cell clumps form typically in shake flasks, which hinders cell growth and decreases protein yield. To minimize clumps during the culture of Chinese hamster ovary cells, we employed the combination of two reagents, dextran sulfate (DS) and recombinant trypsin (r-trypsin). Our results showed that both DS and r-trypsin could diminish cell aggregation when adding them respectively, but clumps were still noticed obviously. In order to further mitigate cell agglomerate, a combination of 1.2 g/L DS and 8.0 mg/L r-trypsin was employed and no clumps were found under the bright field microscope. Strikingly, the highest viable cell density of combination group was increased from 5.12 × 106 to 7.13 × 106 cells/mL, while the integral of viable cells concentration was raised from 35.13 × 106 to 60.87 × 106 cells·days/mL, and the culture period was prolonged by 4 days. In addition, the antibody integrity was maintained in the combination group compared with that of the control.
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Acknowledgments
This research was partly funded by National Natural Science Foundation of China, Ministry of Science and Technology of China (973 and 863 program projects), National Key project for New Drug Development and Manufacture, and Shanghai Commission of Science and Technology as well as special grants from Ministry of Education of China and Shanghai Commission of Education for excellent research team in Oncology and Shanghai Key Project in Oncology.
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Yu Jing and Cunchao Zhang have contributed equally to this study.
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Jing, Y., Zhang, C., Fu, T. et al. Combination of dextran sulfate and recombinant trypsin on aggregation of Chinese hamster ovary cells. Cytotechnology 68, 241–248 (2016). https://doi.org/10.1007/s10616-014-9774-4
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DOI: https://doi.org/10.1007/s10616-014-9774-4