Comparative proteomic analysis of three Chinese hamster ovary (CHO) host cells

Xu, Ningning; Ou, Jianfa; Sun, Wanqi; Zhou, Lufang; Hu, Hui; Liu, Margaret

doi:10.1016/j.bej.2017.05.007

Cited by 29 publications

(17 citation statements)

References 46 publications

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“…Previous investigations that used the systems biology tools have studied differences between the cell line productivity[18-22] and the changes in culture conditions, including the temperature shift[23], sodium butyrate treatment[24,25], and hyperosmolarity[26], which enhance the productivity. Previous proteomic studies have also provided rational strategies for the modifications needed to increase productivity, develop cell growth, improve media consumption, and provide constant glycosylation patterns[27-38].…”

Section: Introductionmentioning

confidence: 99%

Ouantitative Proteomic Analysis of Cellular Responses to a Designed Amino Acid Feed in a Monoclonal Antibody Producing Chinese Hamster Ovary Cell Line

Torkashvand

Mahboudi

Vossoughi

et al. 2018

IBJ

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Background:Chinese hamster ovary (CHO) cell line is considered as the most common cell line in the biopharmaceutical industry because of its capability in performing efficient post-translational modifications and producing the recombinant proteins, which are similar to natural human proteins. The optimization of the upstream process via different feed strategies has a great impact on the target molecule expression and yield.Methods:To determine and understand the molecular events beneath the feed effects on the CHO cell, a label-free quantitative proteomic analysis was applied. The proteome changes followed by the addition of a designed amino acid feed to the monoclonal antibody producing CHO cell line culture medium were investigated.Results:The glutathione synthesis, the negative regulation of the programmed cell death, proteasomal catabolic process, and the endosomal transport pathway were up-regulated in the group fed with a designed amino acid feed compared to the control group.Conclusion:Our findings could be helpful to identify new targets for metabolic engineering.

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Section: Introductionmentioning

confidence: 99%

Ouantitative Proteomic Analysis of Cellular Responses to a Designed Amino Acid Feed in a Monoclonal Antibody Producing Chinese Hamster Ovary Cell Line

Torkashvand

Mahboudi

Vossoughi

et al. 2018

IBJ

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“…Comparing the host cell proteomes of CHO K1, CHO S, and CHO/dhFR − cells, Xu et al . 27 detected 1,307 proteins that were expressed in all three cell lines. Park et al .…”

Section: Discussionmentioning

confidence: 99%

Untargeted proteomics reveals upregulation of stress response pathways during CHO-based monoclonal antibody manufacturing process leading to disulfide bond reduction

Park

Egan

Cura

et al. 2021

mAbs

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Monoclonal antibody (mAb) interchain disulfide bond reduction can cause a loss of function and negatively impact the therapeutic’s efficacy and safety. Disulfide bond reduction has been observed at various stages during the manufacturing process, including processing of the harvested material. The factors and mechanisms driving this phenomenon are not fully understood. In this study, we examined the host cell proteome as a potential factor affecting the susceptibility of a mAb to disulfide bond reduction in the harvested cell culture fluid (HCCF). We used untargeted liquid-chromatography-mass spectrometry-based proteomics experiments in conjunction with a semi-automated protein identification workflow to systematically compare Chinese hamster ovary (CHO) cell protein abundances between bioreactor conditions that result in reduction-susceptible and reduction-free HCCF. Although the growth profiles and antibody titers of these two bioreactor conditions were indistinguishable, we observed broad differences in host cell protein (HCP) expression. We found significant differences in the abundance of glycolytic enzymes, key protein reductases, and antioxidant defense enzymes. Multivariate analysis of the proteomics data determined that upregulation of stress-inducible endoplasmic reticulum (ER) and other chaperone proteins is a discriminatory characteristic of reduction-susceptible HCP profiles. Overall, these results suggest that stress response pathways activated during bioreactor culture increase the reduction-susceptibility of HCCF. Consequently, these pathways could be valuable targets for optimizing culture conditions to improve protein quality.

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“…The established procedure of fed-batch cell culture in 2-L stirred-tank bioreactor for mAb production was described in our previous publication [ 16 ]. The mAb production cultures were seeded with viable cell density (VCD) of 0.3–0.5×10 6 cells/mL in Dynamis medium supplemented with 6 g/L glucose and 8 mM glutamine.…”

Section: Methodsmentioning

confidence: 99%

Bioprocess development of antibody-drug conjugate production for cancer treatment

Goh

et al. 2018

PLoS ONE

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Antibody-drug conjugate (ADC) is a class of targeted cancer therapies that combine the advantages of monoclonal antibody (mAb)’s specific targeting and chemotherapy’s potent cytotoxicity. The therapeutic effect of ADC is significantly affected by its bioproduction process. This study aims to develop an effective ADC production process using anti-HER2 mAb-drug as a model therapeutic. First, a high titer (>2 g/L) of mAb was produced by Chinese hamster ovary cells from fed-batch cell culture. Both live-cell confocal microscopy imaging and flow cytometry analysis demonstrated that the produced mAb and ADC had strong and specific binding to HER2+ cell line BT474. Second, various conjugation conditions of mAb and drug, including linker selection, ratio of drug and mAb, and conjugation approaches, were investigated to improve the production yield and product quality. Finally, the ADC structure and biological quality were evaluated by SDS-PAGE and anti-breast cancer toxicity study, respectively. The ADC with integral molecular structure and high cytotoxicity (IC50 of 1.95 nM) was produced using the optimized production process. The robust bioproduction process could guide the development of ADC-based biopharmaceuticals.

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Comparative proteomic analysis of three Chinese hamster ovary (CHO) host cells

Cited by 29 publications

References 46 publications

Ouantitative Proteomic Analysis of Cellular Responses to a Designed Amino Acid Feed in a Monoclonal Antibody Producing Chinese Hamster Ovary Cell Line

Ouantitative Proteomic Analysis of Cellular Responses to a Designed Amino Acid Feed in a Monoclonal Antibody Producing Chinese Hamster Ovary Cell Line

Untargeted proteomics reveals upregulation of stress response pathways during CHO-based monoclonal antibody manufacturing process leading to disulfide bond reduction

Bioprocess development of antibody-drug conjugate production for cancer treatment

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