Effect of the disulfide isomerase PDIa4 on the antibody production of Chinese hamster ovary cells

Komatsu, Kei; Kumon, Kento; Arita, Mayuno; Onitsuka, Masayoshi; Ômasa, Takeshi; Yohda, Masafumi

doi:10.1016/j.jbiosc.2020.08.001

Cited by 9 publications

(6 citation statements)

References 28 publications

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“…Pdia4, a highly abundant ER protein, plays a critical role in protein folding and assembly (Ferrari and Soling 1999 ), is involved in the response to ER stress (Dorner et al 1990 ), and participates in the formation and isomerization of disulfide bridges (Maattanen et al 2010 ). Contrary to other research findings (Komatsu et al 2020 ), in our data, we did not observe an increase in Pdia4 protein levels upon TM treatment. This observation provides additional support for the hypothesis that the events within the UPR machinery components constitute an early response cascade preceding the full UPR activation, as previously reported (Merksamer et al 2008 ).…”

Section: Discussioncontrasting

confidence: 99%

Quantitative proteomics reveals cellular responses to individual mAb expression and tunicamycin in CHO cells

Sulaj,

Schwaigerlehner,

Sandell

et al. 2024

Appl Microbiol Biotechnol

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Chinese hamster ovary (CHO) cells are popular in the pharmaceutical industry for their ability to produce high concentrations of antibodies and their resemblance to human cells in terms of protein glycosylation patterns. Current data indicate the relevance of CHO cells in the biopharmaceutical industry, with a high number of product commendations and a significant market share for monoclonal antibodies. To enhance the production capabilities of CHO cells, a deep understanding of their cellular and molecular composition is crucial. Genome sequencing and proteomic analysis have provided valuable insights into the impact of the bioprocessing conditions, productivity, and product quality. In our investigation, we conducted a comparative analysis of proteomic profiles in high and low monoclonal antibody–producing cell lines and studied the impact of tunicamycin (TM)-induced endoplasmic reticulum (ER) stress. We examined the expression levels of different proteins including unfolded protein response (UPR) target genes by using label-free quantification techniques for protein abundance. Our results show the upregulation of proteins associated with protein folding mechanisms in low producer vs. high producer cell line suggesting a form of ER stress related to specific protein production. Further, Hspa9 and Dnaja3 are notable candidates activated by the mitochondria UPR and play important roles in protein folding processes in mitochondria. We identified significant upregulation of Nedd8 and Lgmn proteins in similar levels which may contribute to UPR stress. Interestingly, the downregulation of Hspa5/Bip and Pdia4 in response to tunicamycin treatment suggests a low-level UPR activation. Key points • Proteome profiling of recombinant CHO cells under mild TM treatment. • Identified protein clusters are associated with the unfolded protein response (UPR). • The compared cell lines revealed noticeable disparities in protein expression levels.

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Section: Discussioncontrasting

confidence: 99%

Quantitative proteomics reveals cellular responses to individual mAb expression and tunicamycin in CHO cells

Sulaj,

Schwaigerlehner,

Sandell

et al. 2024

Appl Microbiol Biotechnol

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“…Previous studies have shown that PDIA3 and PDIA4 are involved in disulfide bond formation of antibodies in CHO cells. [51] These omics results on the stress response to cystine addition are generally consistent with the results of Ali et al [5,12] Ali et al confirmed an increase in SOD2 with Cys addition, but not an increase in SOD1. [5,12] SOD1 is the predominant SOD in most cells, accounting for 70%-80% of total cellular SOD activity, and is found to be expressed in the cytoplasm.…”

Section: Discussionsupporting

confidence: 87%

“…Previous studies have shown that PDIA3 and PDIA4 are involved in disulfide bond formation of antibodies in CHO cells. [ 51 ]…”

Section: Discussionmentioning

confidence: 99%

Cystine and tyrosine feed reduces oxidative and ER stress in CHO cells

Shibafuji

Nagao

Yohda

2023

Biotechnology Journal

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Multi‐omics analyses was performed to compare the conditions of adding Tyr and Cystine in CHO cells. The addition of cystine resulted in decreased viability and productivity owing to endoplasmic reticulum (ER) stress and the promotion of ER‐associated degradation (ERAD) and apoptosis. In contrast, addition of Tyr suppressed ER stress and apoptosis. This effect could be due to the increase in ubiquinone (Coenzyme Q10) biosynthesized from Tyr. To inhibit apoptosis caused by cystine addition, Tyr was added simultaneously with cystine, which improved growth, viability, and mAb productivity owing to the activation of GSH metabolism, suppression of ER stress and oxidative stress, reduction of ERAD, and activation of the tricarboxylic acid cycle.

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“…In CHO cells, HSP90B1 is induced by glucose limitation or ER stress. 43 , 44 Expression of HSPA9 in CHO cells is increased with accumulation of unfolded protein in the ER. 45 Recently, Xie 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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Effect of the disulfide isomerase PDIa4 on the antibody production of Chinese hamster ovary cells

Cited by 9 publications

References 28 publications

Quantitative proteomics reveals cellular responses to individual mAb expression and tunicamycin in CHO cells

Quantitative proteomics reveals cellular responses to individual mAb expression and tunicamycin in CHO cells

Cystine and tyrosine feed reduces oxidative and ER stress in CHO cells

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

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