| Literature DB >> 25087075 |
Yu Jing1,2,3,4, Cunchao Zhang1,2,3, Tuo Fu1,2,3, Cheng Jiang1,2,3, Kai Ma1,2,3, Dapeng Zhang1,2,3, Sheng Hou1,2,3, Jianxin Dai1,2,3, Hao Wang1,2,3, Xueguang Zhang4, Geng Kou5,6,7, Yajun Guo8,9,10,11.
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 × 10(6) to 7.13 × 10(6) cells/mL, while the integral of viable cells concentration was raised from 35.13 × 10(6) to 60.87 × 10(6) 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.Entities:
Keywords: Aggregation; Chinese hamster ovary cells; Dextran sulfate; Fed-batch; Recombinant trypsin
Year: 2014 PMID: 25087075 PMCID: PMC4754253 DOI: 10.1007/s10616-014-9774-4
Source DB: PubMed Journal: Cytotechnology ISSN: 0920-9069 Impact factor: 2.058