title: Combination of dextran sulfate and recombinant trypsin on aggregation of Chinese hamster ovary cells.
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.
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.|Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Jing
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Cunchao
authors.initials: C
authors.lastname: Zhang
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Tuo
authors.initials: T
authors.lastname: Fu
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Jiang
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Ma
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Dapeng
authors.initials: D
authors.lastname: Zhang
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Hou
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Dai
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China.
authors.forename: Xueguang
authors.initials: X
authors.lastname: Zhang
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China. kou_geng@163.com.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China. kou_geng@163.com.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China. kou_geng@163.com.
authors.forename: Geng
authors.initials: G
authors.lastname: Kou
authors.affiliation: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China. yjguo@smmu.edu.cn.|State Key Laboratory of Antibody Medicine and Target Therapy, Shanghai, 201203, China. yjguo@smmu.edu.cn.|National Engineering Research Center for Antibody Medicine and Shanghai Key Laboratory of Cell Engineering and Antibody, Shanghai, 201203, China. yjguo@smmu.edu.cn.|Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China. yjguo@smmu.edu.cn.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Guo
keywords: Aggregation; Chinese hamster ovary cells; Dextran sulfate; Fed-batch; Recombinant trypsin
references.citation: J Membr Biol. 1973;13(2):97-128
references.pmid: 4205052
references.citation: Biotechnol Bioeng. 2009 Jun 15;103(3):592-608
references.pmid: 19241388
references.citation: Biotechnol Bioeng. 1997 May 5;54(3):191-205
references.pmid: 18634086
references.citation: Biotechnol Prog. 2000 May-Jun;16(3):319-25
references.pmid: 10835230
references.citation: J Virol. 2002 Aug;76(15):7430-43
references.pmid: 12097555
references.citation: J Cell Sci. 1975 Dec;19(3):653-7
references.pmid: 173727
references.citation: Nat Biotechnol. 2004 Nov;22(11):1393-8
references.pmid: 15529164
references.citation: Biotechnol Bioeng. 2013 Feb;110(2):483-90
references.pmid: 23007466
references.citation: Biochem Eng J. 2001 Mar;7(2):121-125
references.pmid: 11173300
references.citation: Mol Biotechnol. 2011 Jan;47(1):9-17
references.pmid: 20589456
references.citation: In Vitro Cell Dev Biol Anim. 2008 Jul-Aug;44(7):179-84
references.pmid: 18594938
references.citation: Anticancer Res. 2005 Mar-Apr;25(2A):895-902
references.pmid: 15868925