|
|
|
| Enhancement of Anti-CD20 Monoclonal Antibody Expression by CHO based on DOE and Amino Acid Supplemental Strategy |
KONG Jian-tao1,ZHUANG Ying-ping1,2,GUO Mei-jin1,2,**( ) |
1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
2 Shanghai Biological Manufacturing Technology Innovation Center, Shanghai 200237, China |
|
|
|
Abstract Fed-batch culture has been one of main bioprocesses for monoclonal antibody (mAb) production by Chinese hamster ovary (CHO) cells. It has been reported that environmental parameters like temperature and pH and nutritional ingredients are key factors that can influence cell growth, carbon and/or nitrogen source metabolism and foreign protein expression in CHO cell suspension culture. Objective: Effects of culture process parameters (temperature and pH) on cell growth and anti-CD20 antibody expression by CHO cells were investigated based on design of experiment (DOE), and a fed-batch strategy was successfully developed to improve anti-CD20 antibody expression level with amino acid supplementation. The results show that temperature was a key factor for anti-CD20 antibody expression: 35℃was the optimal temperature with an increased cell density and target mAb yield. However, the impact of pH on mAb production by CHO cells was not significant and there were no interaction between pH and temperature. The optimal culture conditions were 35℃ and pH7.0 according to the analysis of DOE predictive profiler. In addition, it found that residual concentrations of tyrosine and cysteine in culture was below 0.1mmol/L at late stage of cultivation process under the optimal culture condition. As such, additional 1.5mmol/L tyrosine and 1mmol/L cysteine were fed on day 2, led to an increase in anti-CD20 antibody titer by 24.1% and no changes in glycosylation of anti-CD20 antibody.
|
|
Received: 06 September 2020
Published: 14 January 2021
|
|
|
|
Corresponding Authors:
Mei-jin GUO
E-mail: guo_mj@ecust.edu.cn
|
|
|
| [1] |
Werner R G, Noe W, Kopp K, et al. Appropriate mammalian expression systems for biopharmaceuticals. Arzneimittel-Forschung, 1998,48(8):870-880.
pmid: 9748718
|
|
|
| [2] |
Raju T S. Terminal sugars of Fc glycans influence antibody effector functions of IgGs. Current Opinion in Immunology, 2008,20(4):471-478.
doi: 10.1016/j.coi.2008.06.007
pmid: 18606225
|
|
|
| [3] |
Huang Y M, Hu W W, Rustandi E, et al. Maximizing productivity of CHO cell-based fed-batch culture using chemically defined media conditions and typical manufacturing equipment. Biotechnology Progress, 2010,26(5):1400-1410.
pmid: 20945494
|
|
|
| [4] |
Bollati-Fogolin M, Forno G, Nimtz M, et al. Temperature reduction in cultures of hGM-CSF-expressing CHO cells:Effect on productivity and product quality. Biotechnology Progress, 2005,21(1):17-21.
pmid: 15903236
|
|
|
| [5] |
Yoon S K, Choi S L, Song J Y, et al. Effect of culture pH on erythropoietin production by Chinese Hamster ovary cells grown in suspension at 32.5 and 37.0℃. Biotechnology and Bioengineering, 2005,89(3):345-356.
pmid: 15625678
|
|
|
| [6] |
Jardon M, Garnier A. pH, pCO2, and temperature effect on R-Adenovirus production. Biotechnology Progress, 2003,19(1):202-208.
doi: 10.1021/bp025585a
pmid: 12573026
|
|
|
| [7] |
Kaufmann H, Mazur X, Fussenegger M, et al. Influence of low temperature on productivity, proteome and protein phosphorylation of CHO cells. Biotechnology and Bioengineering, 1999,63(5):573-582.
doi: 10.1002/(sici)1097-0290(19990605)63:5<573::aid-bit7>3.0.co;2-y
pmid: 10397813
|
|
|
| [8] |
Yoon S K, Song J Y, Lee G M. Effect of low culture temperature on specific productivity, transcription level, and heterogeneity of erythropoietin in Chinese hamster ovary cells. Biotechnology and Bioengineering, 2003,82(3):289-298.
doi: 10.1002/bit.10566
pmid: 12599255
|
|
|
| [9] |
Xing Z, Kenty B, Koyrakh I, et al. Optimizing amino acid composition of CHO cell culture media for a fusion protein production. Process Biochemistry, 2011,46(7):1423-1429.
|
|
|
| [10] |
Kang S, Mullen J, Miranda L P, et al. Utilization of tyrosine-and histidine-containing dipeptides to enhance productivity and culture viability. Biotechnology and Bioengineering, 2012,109(9):2286-2294.
doi: 10.1002/bit.24507
pmid: 22447498
|
|
|
| [11] |
Xing Z Z, Li Z J, Chow V, et al. Identifying inhibitory threshold values of repressing metabolites in CHO cell culture using multivariate analysis methods. Biotechnology Progress, 2008,24(3):675-683.
pmid: 18422365
|
|
|
| [12] |
Woolley J F, Al-Rubeai M. The application of SELDI-TOF mass spectrometry to mammalian cell culture. Biotechnology Advances, 2009,27(2):177-184.
pmid: 19049820
|
|
|
| [13] |
Li F, Zhou J X, Yang X, et al. Current therapeutic antibody production and process optimization. Bioprocessing Journal, 2006,5(4):16-25.
doi: 10.12665/issn.1538-8786
|
|
|
| [14] |
Feeney L, Carvalhal V, Yu X C, et al. Eliminating tyrosine sequence variants in CHO cell lines producing recombinant monoclonal antibodies. Biotechnology and Bioengineering, 2013,110(4):1087-1097.
doi: 10.1002/bit.24759
|
|
|
| [15] |
庄少颖, 史劲松. 重组抗CD20单克隆抗体细胞培养工艺优化. 名医, 2020,12:349-352.
|
|
|
| [15] |
Zhuang S Y, Shi J S. The optimization of cell culture process for recombinant anti-CD20 monoclonal antibody. Renowned Doctor, 2020,12:349-352.
|
|
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
