Reduction of charge variants by CHO cell culture process optimization

Weng, Zhibing; Jin, Jian; Shao, ChunHua; Li, Huazhong

doi:10.1007/s10616-020-00375-x

Cited by 21 publications

(19 citation statements)

References 56 publications

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“…In a chemostat, a cell culture is maintained in a continuous regime where fresh medium is pumped into the culture vessel at the same rate that it is extracted, such that the working volume remains constant [44]. In Figure (1) we present a schematic picture of the chemostat. The dynamics of cell and metabolite concentrations in the vessel, X (gCDW × l −1 ) and s i (mM ), for a classic Adapted from [17].…”

Section: B Constraint-based Modeling Of the Chemostatmentioning

confidence: 99%

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Inferring metabolic fluxes in nutrient-limited continuous cultures: A Maximum Entropy Approach with minimum information

A.¹,

Fernández-de-Cossio-Díaz²,

Mulet³

2021

Preprint

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We propose a new scheme to infer the metabolic fluxes of cell cultures in a chemostat.Our approach is based on the Maximum Entropy Principle and exploits the understanding of the chemostat dynamics and its connection with the actual metabolism of cells. We show that, in continuous cultures with limiting nutrients, the inference can be done with limited information about the culture: the dilution rate of the chemostat, the concentration in the feed media of the limiting nutrient and the cell concentration at steady state. Also, we remark that our technique provides information, not only about the mean values of the fluxes in the culture, but also its heterogeneity. We first present these results studying a computational model of a chemostat. Having control of this model we can test precisely the quality of the inference, and also unveil the mechanisms behind the success of our approach.Then, we apply our method to E. coli experimental data from the literature and show that it outperforms alternative formulations that rest on a Flux Balance Analysis framework.

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Section: B Constraint-based Modeling Of the Chemostatmentioning

confidence: 99%

“…The study of cellular metabolism is a research field with a direct impact on the biotechnological industry. Indeed, cell culture-derived products are a major part of a multi-billion market [1]. These products are obtained by exploiting the capabilities of cellular metabolism to produce molecules with a wide range of chemical complexity.…”

Section: Introductionmentioning

confidence: 99%

Inferring metabolic fluxes in nutrient-limited continuous cultures: A Maximum Entropy Approach with minimum information

A.¹,

Fernández-de-Cossio-Díaz²,

Mulet³

2021

Preprint

View full text Add to dashboard Cite

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“…In 1986, the first recombinant therapeutic protein, tissue plasminogen activator (tPA), was approved for marketing ( Kaufman et al, 1985 ), its expression titer was less than 50 mg/L. After 30 years of rapidly development and technological breakthroughs, progress in therapeutic antibody field has been accelerated, the antibody expression titers have increased more than 100–200 times, even the yield of therapeutic antibody has exceeded 10 g/L ( Li et al, 2010 ; Weng et al, 2020 ). The approved therapeutic monoclonal antibodies produced in CHO cells until 2020 are listed in Table 1 .…”

Section: Introductionmentioning

confidence: 99%

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Zhang

Shan

Liang

et al. 2022

Front. Bioeng. Biotechnol.

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Recombinant antibodies are rapidly developing therapeutic agents; approximately 40 novel antibody molecules enter clinical trials each year, most of which are produced from Chinese hamster ovary (CHO) cells. However, one of the major bottlenecks restricting the development of antibody drugs is how to perform high-level expression and production of recombinant antibodies. The high-efficiency expression and quality of recombinant antibodies in CHO cells is determined by multiple factors. This review provides a comprehensive overview of several state-of-the-art approaches, such as optimization of gene sequence of antibody, construction and optimization of high-efficiency expression vector, using antibody expression system, transformation of host cell lines, and glycosylation modification. Finally, the authors discuss the potential of large-scale production of recombinant antibodies and development of culture processes for biopharmaceutical manufacturing in the future.

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“…However, it was also shown that an improved energy metabolism under hyperosmotic conditions favors specific productivity of CHO cells (Lee et al, 2003). While most of these studies focused on the effects of one factor at a time, more systemic approaches such as quality by design and design of experiments have been used to study the link between applied parameters whilst further pushing the limits of recombinant protein production (Looby et al, 2011;Nagashima et al, 2013;Weng et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Proteomic Profiling of IgG1 Producing CHO Cells Using LC/LC-SPS-MS3: The Effects of Bioprocessing Conditions on Productivity and Product Quality

Strasser

Farrell

et al. 2021

Front. Bioeng. Biotechnol.

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The biopharmaceutical market is dominated by monoclonal antibodies, the majority of which are produced in Chinese hamster ovary (CHO) cell lines. Intense cell engineering, in combination with optimization of various process parameters results in increasing product titers. To enable further improvements in manufacturing processes, detailed information about how certain parameters affect cellular mechanisms in the production cells, and thereby also the expressed drug substance, is required. Therefore, in this study the effects of commonly applied changes in bioprocessing parameters on an anti-IL8 IgG1 producing CHO DP-12 cell line were investigated on the level of host cell proteome expression combined with product quality assessment of the expressed IgG1 monoclonal antibody. Applying shifts in temperature, pH and dissolved oxygen concentration, respectively, resulted in altered productivity and product quality. Furthermore, analysis of the cells using two-dimensional liquid chromatography-mass spectrometry employing tandem mass tag based isotopic quantitation and synchronous precursor selection-MS3 detection revealed substantial changes in the protein expression profiles of CHO cells. Pathway analysis indicated that applied bioprocessing conditions resulted in differential activation of oxidative phosphorylation. Additionally, activation of ERK5 and TNFR1 signaling suggested an affected cell cycle. Moreover, in-depth product characterization by means of charge variant analysis, peptide mapping, as well as structural and functional analysis, revealed posttranslational and structural changes in the expressed drug substance. Taken together, the present study allows the conclusion that, in anti-IL8 IgG1 producing CHO DP-12 cells, an improved energy metabolism achieved by lowering the cell culture pH is favorable when aiming towards high antibody production rates while maintaining product quality.

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Reduction of charge variants by CHO cell culture process optimization

Cited by 21 publications

References 56 publications

Inferring metabolic fluxes in nutrient-limited continuous cultures: A Maximum Entropy Approach with minimum information

Inferring metabolic fluxes in nutrient-limited continuous cultures: A Maximum Entropy Approach with minimum information

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Proteomic Profiling of IgG1 Producing CHO Cells Using LC/LC-SPS-MS3: The Effects of Bioprocessing Conditions on Productivity and Product Quality

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