scholarly journals Designing a Strategy for pH Control to Improve CHO Cell Productivity in Bioreactor

2021 ◽  
Author(s):  
Zohreh Ahleboot ◽  
Mahdi Khorshidtalab ◽  
Paria Motahari ◽  
Rasoul Mahboudi ◽  
Razieh Arjmand ◽  
...  
Keyword(s):  
Cell Culture ◽  
Large Scale ◽  
Chinese Hamster ◽  
Ph Control ◽  
Agitation Speed ◽  
Cell Culture Process ◽  
Co2 Removal ◽  
Bioreactor Culture ◽  
Product Titer ◽  
Culture Ph

Background: Drastic pH drop is a common consequence of scaling up a mammalian cell culture process, where it may affect the final performance of cell culture. Although CO2 sparging and base addition are used as common approaches for pH control, these strategies are not necessarily successful in large scale bioreactors due to their effect on osmolality and cell viability. Accordingly, a series of experiments were conducted using an IgG1 producing Chinese Hamster Ovary (CHO-S) cell culture in 30 L bioreactor to assess the efficiency of an alternative strategy in controlling culture pH.   Methods: Factors inducing partial pressure of CO2 and lactate accumulation (as the main factors altering culture pH) were assessed by Plackett-Burman design to identify the significant ones. As culture pH directly influences process productivity, protein titer was measured as the response variable. Subsequently, Central Composite Design (CCD) was employed to obtain a model for product titer prediction as a function of individual and interaction effects of significant variables.   Results: The results indicated that the major factor affecting pH is non-efficient CO2 removal. CO2 accumulation was found to be affected by an interaction between agitation speed and overlay air flow rate. Accordingly, after increasing the agitation speed and headspace aeration, the culture pH was successfully maintained in the range of 6.95-7.1, resulting in 51% increase in final product titer. Similar results were obtained during 250 L scale bioreactor culture, indicating the scalability of the approach.   Conclusion: The obtained results showed that pH fluctuations could be effectively controlled by optimizing CO2 stripping.

Scale-up analysis for a CHO cell culture process in large-scale bioreactors

2009 ◽  
Vol 103 (4) ◽  
pp. 733-746 ◽  
Author(s):  
Zizhuo Xing ◽  
Brian M. Kenty ◽  
Zheng Jian Li ◽  
Steven S. Lee
Keyword(s):  
Cell Culture ◽  
Large Scale ◽  
Scale Up ◽  
Cell Culture Process ◽  
Cho Cell ◽  
Culture Process ◽  
Cho Cell Culture

scholarly journals Cell-culture growth conditions resulting in the oxidation of a recombinant antigen-binding fragment

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Khandaker Siddiquee ◽  
Cheng Zhao ◽  
Michelle A. Stemler ◽  
Bill Zeck ◽  
Jeffrey R. Fishpaugh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Feeding Strategy ◽  
Scale Up ◽  
Chinese Hamster ◽  
Recombinant Antigen ◽  
Small Scale ◽  
Cell Culture Process ◽  
Antigen Binding ◽  
Scale Production ◽  
Target Molecule

Abstract Use of Quality-by-Design (QbD) tools is becoming an important part of the bioprocessing industry when developing a process for manufacturing operations to ensure the robustness and reproducibility of the biologic product. In the present study, a QbD tool, Design of Experiments (DOE), was utilized to optimize a bioprocess for the production of a CHO recombinant antigen-binding fragment (rFab) in small-scale bioreactors. DOE studies evaluated percent dissolved oxygen, temperature, and feeding strategy specific to this Chinese Hamster Ovary (CHO) clone. It was determined that these factors influenced cell viability, yield of the recombinant protein, and metabolic byproduct formation. To ensure the quality of the target molecule in the cell-culture process, small-scale purifications and analytical evaluation of the target molecule were completed prior to cell-culture scale-up to ensure that oxidation of the rFab, presence of free light chain, and truncation of thiol group were not observed. Analysis of the purified rFab by mass spectrometry indicated that rFab oxidation occurred under poor cell-culture conditions. PCR profile array results also revealed increased transcription of the oxidative genes Superoxide Dismutase 3, Myeloperoxidase, Dual Oxidase Like 2, Nuclear Receptor Coactivator 7, NADPH Oxidase Organizer 1, Mitochondria Uncouple Protein 3, Eosinophil Peroxidase, Lactoperoxidase Like, Serum Albumin Like, and Glutathione S-Transferase Pi 1 in this CHO strain. The present study suggests a mechanism and pathway for the oxidation of an rFab molecule during cell-culture bioprocess optimization. The present study also demonstrated the importance of utilizing the QbD tool of DOE to optimize the cell-culture bioprocess prior to scaling up into the large-scale production bioreactor.

scholarly journals Effect of Sialic Acid on Mammalian Cell Culture and Protein Expression: A Potential Productivity Enhancer for Biopharmaceutical Cell Culture Processes

Processes ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 1449
Author(s):  
Xiangsong Chen ◽  
Shang Xiao ◽  
Jinyong Wu ◽  
Jianming Yao
Keyword(s):  
Cell Culture ◽  
Sialic Acid ◽  
Cell Lines ◽  
Hek293 Cell ◽  
Chinese Hamster ◽  
Lactate Production ◽  
Production Costs ◽  
Cell Culture Process ◽  
Cho Cell ◽  
Potential Productivity

Improved productivity of the two most commonly used cell lines in the biopharmaceutical industry, such as human embryonic kidney 293 (HEK293) and Chinese hamster ovary (CHO), could reduce production costs and increase manufacturing capacity. One method for increasing protein productivity is the addition of antioxidants during the cell culture process. In this study, we examined the effect of sialic acid (SA) on one HEK293 cell line and two CHO cell lines. The addition of SA to HEK293 cell led to a higher viable cell density (VCD), viability (Via), and a lower lactate content in the later stage of cultures. Further results showed that SA reduced the reactive oxygen species (ROS), improved cell viability, reduced lactate production, and increased antibody expression by more than 20% in the later stage of the two CHO cell lines cultures. Besides, an optimized dose of SA had no significant effect on acidic variants level aggregation level, N-linked glycosylation pattern, and SA content on antibodies. These results suggest that the addition of SA can improve the productivity of biopharmaceutical cell culture processes.

scholarly journals CHOmpact: a reduced metabolic model of Chinese hamster ovary cells with enhanced interpretability

2021 ◽  
Author(s):  
Ioscani Jimenez del Val ◽  
Sarantos Kyriakopoulos ◽  
Simone Albrecht ◽  
Henning Stöckmann ◽  
Pauline M Rudd ◽  
...  
Keyword(s):  
Cell Culture ◽  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Large Scale ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Reaction Network ◽  
Metabolic Model ◽  
Feeding Strategies ◽  
Metabolic Models

Metabolic modelling has emerged as a key tool for the characterisation of biopharmaceutical cell culture processes. Metabolic models have also been instrumental in identifying genetic engineering targets and developing feeding strategies that optimise the growth and productivity of Chinese hamster ovary (CHO) cells. Despite their success, metabolic models of CHO cells still present considerable challenges. Genome scale metabolic models (GeMs) of CHO cells are very large (>6000 reactions) and are, therefore, difficult to constrain to yield physiologically consistent flux distributions. The large scale of GeMs also makes interpretation of their outputs difficult. To address these challenges, we have developed CHOmpact, a reduced metabolic network that encompasses 101 metabolites linked through 144 reactions. Our compact reaction network allows us to deploy multi-objective optimisation and ensure that the computed flux distributions are physiologically consistent. Furthermore, our CHOmpact model delivers enhanced interpretability of simulation results and has allowed us to identify the mechanisms governing shifts in the anaplerotic consumption of asparagine and glutamate as well as an important mechanism of ammonia detoxification within mitochondria. CHOmpact, thus, addresses key challenges of large-scale metabolic models and, with further development, will serve as a platform to develop dynamic metabolic models for the control and optimisation of biopharmaceutical cell culture processes.

Comparative metabolite analysis to understand lactate metabolism shift in Chinese hamster ovary cell culture process

2011 ◽  
Vol 109 (1) ◽  
pp. 146-156 ◽  
Author(s):  
Jun Luo ◽  
Natarajan Vijayasankaran ◽  
Jennifer Autsen ◽  
Rodell Santuray ◽  
Terry Hudson ◽  
...  
Keyword(s):  
Cell Culture ◽  
Chinese Hamster Ovary Cell ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Cell Culture Process ◽  
Lactate Metabolism ◽  
Metabolite Analysis ◽  
Culture Process

scholarly journals High shear resistance of insect cells: the basis for substantial improvements in cell culture process design

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Florian Strobl ◽  
Mark Duerkop ◽  
Dieter Palmberger ◽  
Gerald Striedner
Keyword(s):  
Cell Culture ◽  
Gas Flow ◽  
Insect Cells ◽  
Negative Impact ◽  
Chinese Hamster ◽  
Shear Resistance ◽  
Pure Oxygen ◽  
Cell Culture Process ◽  
Foam Formation ◽  
Shear Rates

AbstractMulticellular organisms cultivated in continuous stirred tank reactors (CSTRs) are more sensitive to environmental conditions in the suspension culture than microbial cells. The hypothesis, that stirring induced shear stress is the main problem, persists, although it has been shown that these cells are not so sensitive to shear. As these results are largely based on Chinese Hamster Ovary (CHO) cell experiments the question remains if similar behavior is valid for insect cells with a higher specific oxygen demand. The requirement of higher oxygen transfer rates is associated with higher shear forces in the process. Consequently, we focused on the shear resistance of insect cells, using CHO cells as reference system. We applied a microfluidic device that allowed defined variations in shear rates. Both cell lines displayed high resistance to shear rates up to 8.73 × 105 s−1. Based on these results we used microbial CSTRs, operated at high revolution speeds and low aeration rates and found no negative impact on cell viability. Further, this cultivation approach led to substantially reduced gas flow rates, gas bubble and foam formation, while addition of pure oxygen was no longer necessary. Therefore, this study contributes to the development of more robust insect cell culture processes.

scholarly journals Mechanistic understanding of CHO cell culture improvement by rosmarinic acid through multi-omics analysis

2021 ◽  
Author(s):  
Zhuangrong Huang ◽  
Jianlin Xu ◽  
Jun Tian ◽  
Kathryn Aron ◽  
Yueming Qian ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Rosmarinic Acid ◽  
Cell Biology ◽  
Chinese Hamster ◽  
Viable Cell ◽  
Cell Culture Process ◽  
Cho Cell ◽  
Critical Pathways ◽  
Omics Analysis

The use of antioxidants in Chinese hamster ovary (CHO) cell cultures to improve monoclonal antibody production has been a topic of great interest. Nevertheless, the mechanisms by which antioxidant pathways are regulated in CHO cells and their effect on metabolism are not fully understood. In this work, we investigated how treatment with the antioxidant rosmarinic acid (RA) improved viable cell density and titer in CHO cell cultures, and attempted to explore the underlying mechanism(s) using transcriptomics and metabolomics. In particular, transcriptomics analysis indicated that RA treatment modified gene expression and strongly affected the MAPK and Akt signaling pathways which regulate cell survival and cell death. Moreover, we observed that these effects did not appear related to an intracellular metabolism change. In summary, this integrated ‘omics analysis has important implications for the role of the antioxidant RA in industrial cell culture processes. The current study also represents an example in the industry of how multi-omics can be applied to gain an in‐depth understanding of CHO cell biology and to identify critical pathways that can contribute to cell culture process improvement and cell line engineering.

Characterizing the preparation of a concentrated nutrient feed solution for a large-scale cell culture process

2018 ◽  
Vol 134 ◽  
pp. 120-128
Author(s):  
Brian Russell ◽  
Guillermo Miro-Quesada ◽  
Qu Limin ◽  
Sanjeev Ahuja
Keyword(s):  
Cell Culture ◽  
Large Scale ◽  
Feed Solution ◽  
Cell Culture Process ◽  
Culture Process ◽  
Nutrient Feed

Miniature Bioreactors for Rapid Bioprocess Development of Mammalian Cell Culture

2012 ◽  
Vol 59 (1) ◽  
Author(s):  
Mohd Helmi Sani ◽  
Frank Baganz
Keyword(s):  
Cell Culture ◽  
Mammalian Cell ◽  
Process Development ◽  
Mammalian Cell Culture ◽  
Small Scale ◽  
Cell Culture Process ◽  
Deep Wells ◽  
Working Volume ◽  
Culture Process ◽  
And Control

At present, there are a number of commercial small scale shaken systems available on the market with instrumented controllable microbioreactors such as Micro–24 Microreactor System (Pall Corporation, Port Washington, NY) and M2P Biolector, (M2P Labs GmbH, Aachen, Germany). The Micro–24 system is basically an orbital shaken 24–well plate that operates at working volume 3 – 7 mL with 24 independent reactors (deep wells, shaken and sparged) running simultaneously. Each reactor is designed as single use reactor that has the ability to continuously monitor and control the pH, DO and temperature. The reactor aeration is supplied by sparging air from gas feeds that can be controlled individually. Furthermore, pH can be controlled by gas sparging using either dilute ammonia or carbon dioxide directly into the culture medium through a membrane at the bottom of each reactor. Chen et al., (2009) evaluated the Micro–24 system for the mammalian cell culture process development and found the Micro–24 system is suitable as scaledown tool for cell culture application. The result showed that intra-well reproducibility, cell growth, metabolites profiles and protein titres were scalable with 2 L bioreactors.

scholarly journals Modelling and pH Control in Raceway and Thin-Layer Photobioreactors for Wastewater Treatment

Energies ◽  
2021 ◽  
Vol 14 (4) ◽  
pp. 1099
Author(s):  
María José Rodríguez-Torres ◽  
Ainoa Morillas-España ◽  
José Luis Guzmán ◽  
Francisco Gabriel Acién
Keyword(s):  
Wastewater Treatment ◽  
Thin Layer ◽  
Large Scale ◽  
Control Period ◽  
Ph Control ◽  
Thin Layers ◽  
Control Algorithms ◽  
Treatment Processes ◽  
Overall Performance ◽  
Reduction Of Co2

One of the most critical variables in microalgae-related processes is the pH; it directly determines the overall performance of the production system especially when coupling with wastewater treatment. In microalgae-related wastewater treatment processes, the adequacy of pH has a large impact on the microalgae/bacteria consortium already developing on these systems. For cost-saving reasons, the pH is usually controlled by classical On/Off control algorithms during the daytime period, typically with the dynamics of the system and disturbances not being considered in the design of the control system. This paper presents the modelling and pH control in open photobioreactors, both raceway and thin-layer, using advanced controllers. In both types of photobioreactors, a classic control was implemented and compared with a Proportional–Integral (PI) control, also the operation during only the daylight period and complete daily time was evaluated. Thus, three major variables already studied include (i) the type of reactors (thin-layers and raceways), (ii) the type of control algorithm (On/Off and PI), and (iii) the control period (during the daytime and throughout the daytime and nighttime). Results show that the pH was adequately controlled in both photobioreactors, although each type requires different control algorithms, the pH control being largely improved when using PI controllers, with the controllers allowing us to reduce the total costs of the process with the reduction of CO2 injections. Moreover, the control during the complete daily cycle (including night) not only not increases the amount of CO2 to be injected, otherwise reducing it, but also improves the overall performance of the production process. Optimal pH control systems here developed are highly useful to develop robust large-scale microalgae-related wastewater treatment processes.

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