scholarly journals A Metabolomics Approach to Increasing Chinese Hamster Ovary (CHO) Cell Productivity

Metabolites ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 823
Author(s):  
Grace Yao ◽  
Kathryn Aron ◽  
Michael Borys ◽  
Zhengjian Li ◽  
Girish Pendse ◽  
...  
Keyword(s):  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Tca Cycle ◽  
Viable Cell ◽  
Metabolic Model ◽  
Viable Cell Density ◽  
Specific Productivity ◽  
Cho Cell ◽  
High Productivity ◽  
Medium Supplementation

Much progress has been made in improving the viable cell density of bioreactor cultures in monoclonal antibody production from Chinese hamster ovary (CHO) cells; however, specific productivity (qP) has not been increased to the same degree. In this work, we analyzed a library of 24 antibody-expressing CHO cell clones to identify metabolites that positively associate with qP and could be used for clone selection or medium supplementation. An initial library of 12 clones, each producing one of two antibodies, was analyzed using untargeted LC-MS experiments. Metabolic model-based annotation followed by correlation analysis detected 73 metabolites that significantly correlated with growth, qP, or both. Of these, metabolites in the alanine, aspartate, and glutamate metabolism pathway, and the TCA cycle showed the strongest association with qP. To evaluate whether these metabolites could be used as indicators to identify clones with potential for high productivity, we performed targeted LC-MS experiments on a second library of 12 clones expressing a third antibody. These experiments found that aspartate and cystine were positively correlated with qP, confirming the results from untargeted analysis. To investigate whether qP correlated metabolites reflected endogenous metabolic activity beneficial for productivity, several of these metabolites were tested as medium additives during cell culture. Medium supplementation with citrate improved qP by up to 490% and more than doubled the titer. Together, these studies demonstrate the potential for using metabolomics to discover novel metabolite additives that yield higher volumetric productivity in biologics production processes.

scholarly journals FACTORS AFFECTING CHINESE HAMSTER OVARY CELL PROLIFERATION AND VIABILITY

2019 ◽  
Vol 1 ◽  
pp. 245
Author(s):  
Alina Rekena ◽  
Dora Livkisa ◽  
Dagnija Loca
Keyword(s):  
Cell Proliferation ◽  
Cell Line ◽  
Cell Density ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Fed Batch ◽  
Cho Cell ◽  
Decline Phase

Advantageous cultivation procedures for the Chinese hamster ovary (CHO) cells are necessary for the productive commercial production of biopharmaceuticals. A main challenge that needs to be addressed during the process development is the differences in each cell line requirements concerning the nutrients and feed strategies in order to achieve the desired growth characteristics. Therefore, within the current research, a naïve high cell density serum free suspension adapted CHO cell line was tested with glucose and glutamine rich feeds in fed-batch Erlenmeyer shake flask cultures. Glucose consumption rate was adjusted to develop the optimal feed strategies. Obtained results indicated that high glucose and l-glutamine feeding did not improve maximum viable cell density compared to the control samples. During the exponential phase, cell proliferation and viability of all feeds showed no statistically significant difference. Instead, the fed-batch processes tested led to statistically significant differences in viable cell density and cell viability during the decline phase, compared to control (batch) culture. The difference between glucose and glutamine feeding was indistinguishable, most probably due to the concentration imbalance with the rest of the nutrients in feed. The overall study presented a method to slow down the decrease in CHO cell proliferation and viability during the decline phase, instead of increasing the maximum cell density at the plateau phase. 

E2F-1 overexpression increases viable cell density in batch cultures of Chinese hamster ovary cells

2008 ◽  
Vol 138 (3-4) ◽  
pp. 103-106 ◽  
Author(s):  
Brian S. Majors ◽  
Nilou Arden ◽  
George A. Oyler ◽  
Gisela G. Chiang ◽  
Nels E. Pederson ◽  
...  
Keyword(s):  
Cell Density ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Ovary Cells ◽  
Batch Cultures

scholarly journals Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

Antioxidants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 159 ◽  
Author(s):  
Luis Toronjo-Urquiza ◽  
David James ◽  
Tibor Nagy ◽  
Robert Falconer
Keyword(s):  
Cell Culture ◽  
Cell Density ◽  
Culture Media ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Specific Productivity ◽  
Cho Cell ◽  
Cell Culture Media ◽  
Igg Synthesis ◽  
Cho Cell Culture

This study identified several antioxidants that could be used in Chinese hamster ovary (CHO)cell culture media and benefit monoclonal antibody production. The flavan-3-ols, catechin, epicatechin, epigallocatechin gallate and gallocatechin gallate all had no detrimental effect on cell viability at the concentrations tested, and they reduced the final viable cell count with a resulting rise in the cell specific productivity. The flavone, luteolin behave similarly to the flavan-3-ols. Resveratrol at 50 μM concentration resulted in the most pronounced reduction in viable cell density with minimal decrease in IgG synthesis and the largest increase in cell specific productivity. Low concentrations of α-tocopherol (35 μM) reduced viable cell density and raised cell specific productivity, but at higher concentration it had little additional effect. As high concentrations of α-tocopherol are not toxic to CHO cells, its addition as an anti-oxidant has great potential. Kaempferol up to 50 μM, curcumin up to 20 μM and piceid up to 100 μM showed little effect on growth or IgG synthesis and could be useful as antioxidants. Caffeic acid phenethyl ester was toxic to CHO cell and of no interest. Seven of the phenolic compounds tested are potential cell cycle inhibitors as well as having intrinsic antioxidant properties.

scholarly journals Off-Gas-Based Soft Sensor for Real-Time Monitoring of Biomass and Metabolism in Chinese Hamster Ovary Cell Continuous Processes in Single-Use Bioreactors

Processes ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 2073
Author(s):  
Tobias Wallocha ◽  
Oliver Popp
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Chinese Hamster Ovary ◽  
Mammalian Cell Culture ◽  
Chinese Hamster ◽  
Viable Cell ◽  
Soft Sensor ◽  
Cho Cell ◽  
Continuous Processes ◽  
Single Use

In mammalian cell culture, especially in pharmaceutical manufacturing and research, biomass and metabolic monitoring are mandatory for various cell culture process steps to develop and, finally, control bioprocesses. As a common measure for biomass, the viable cell density (VCD) or the viable cell volume (VCV) is widely used. This study highlights, for the first time, the advantages of using VCV instead of VCD as a biomass depiction in combination with an oxygen-uptake- rate (OUR)-based soft sensor for real-time biomass estimation and process control in single-use bioreactor (SUBs) continuous processes with Chinese hamster ovary (CHO) cell lines. We investigated a series of 14 technically similar continuous SUB processes, where the same process conditions but different expressing CHO cell lines were used, with respect to biomass growth and oxygen demand to calibrate our model. In addition, we analyzed the key metabolism of the CHO cells in SUB perfusion processes by exometabolomic approaches, highlighting the importance of cell-specific substrate and metabolite consumption and production rate qS analysis to identify distinct metabolic phases. Cell-specific rates for classical mammalian cell culture key substrates and metabolites in CHO perfusion processes showed a good correlation to qOUR, yet, unexpectedly, not for qGluc. Here, we present the soft-sensoring methodology we developed for qPyr to allow for the real-time approximation of cellular metabolism and usage for subsequent, in-depth process monitoring, characterization and optimization.

scholarly journals Glycosylation Flux Analysis of Immunoglobulin G in Chinese Hamster Ovary Perfusion Cell Culture

Processes ◽  
2018 ◽  
Vol 6 (10) ◽  
pp. 176 ◽  
Author(s):  
Sandro Hutter ◽  
Moritz Wolf ◽  
Nan Papili Gao ◽  
Dario Lepori ◽  
Thea Schweigler ◽  
...  
Keyword(s):  
Cell Culture ◽  
Immunoglobulin G ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Specific Productivity ◽  
Glycan Structure ◽  
Flux Analysis ◽  
Cho Cell ◽  
Clinical Safety ◽  
Perfusion Cell Culture

The terminal sugar molecules of the N-linked glycan attached to the fragment crystalizable (Fc) region is a critical quality attribute of therapeutic monoclonal antibodies (mAbs) such as immunoglobulin G (IgG). There exists naturally-occurring heterogeneity in the N-linked glycan structure of mAbs, and such heterogeneity has a significant influence on the clinical safety and efficacy of mAb drugs. We previously proposed a constraint-based modeling method called glycosylation flux analysis (GFA) to characterize the rates (fluxes) of intracellular glycosylation reactions. One contribution of this work is a significant improvement in the computational efficiency of the GFA, which is beneficial for analyzing large datasets. Another contribution of our study is the analysis of IgG glycosylation in continuous perfusion Chinese Hamster Ovary (CHO) cell cultures. The GFA of the perfusion cell culture data indicated that the dynamical changes of IgG glycan heterogeneity are mostly attributed to alterations in the galactosylation flux activity. By using a random forest regression analysis of the IgG galactosylation flux activity, we were further able to link the dynamics of galactosylation with two process parameters: cell-specific productivity of IgG and extracellular ammonia concentration. The characteristics of IgG galactosylation dynamics agree well with what we previously reported for fed-batch cultivations of the same CHO cell strain.

Review of the current methods of Chinese Hamster Ovary (CHO) cells cultivation for production of therapeutic protein

2020 ◽  
Vol 17 ◽  
Author(s):  
Shazid Md. Sharker ◽  
Md. Atiqur Rahman
Keyword(s):  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Mass Production ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Therapeutic Protein ◽  
Specific Productivity ◽  
Ovary Cells ◽  
Further Development ◽  
Interesting Area

Most of clinical approved protein-based drugs or under in clinical trial have a profound impact in the treatment of critical diseases. The mammalian eukaryotic cells culture approaches, particularly the CHO (Chinese Hamster Ovary) cells are mainly used in the biopharmaceutical industry for the mass-production of therapeutic protein. Recent advances in CHO cell bioprocessing to yield recombinant proteins and monoclonal antibodies have enabled the expression of quality protein. The developments of cell lines are possible to upgrade specific productivity. As a result, it holds an interesting area for academic as well as industrial researchers around the world. This review will concentrate on the recent progress of the mammalian CHO cells culture technology and the future scope of further development for the mass-production of protein therapeutics.

scholarly journals Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Laura Bryan ◽  
Michael Henry ◽  
Ronan M. Kelly ◽  
Christopher C. Frye ◽  
Matthew D. Osborne ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Density ◽  
Cell Lines ◽  
Viable Cell ◽  
Viable Cell Density ◽  
High Peak ◽  
Proteomic Profiling ◽  
Cho Cell ◽  
Extended Culture ◽  
Key Pathways

Abstract Background The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost of goods. In this study we implemented label-free LC-MS/MS proteomic profiling of IgG4 producing CHO cell lines throughout the duration of the cell culture to identify differentially expressed (DE) proteins and intracellular pathways associated with the high peak viable cell density (VCD) and extended culture VCD phenotypes. Results We identified key pathways in DNA replication, mitotic cell cycle and evasion of p53 mediated apoptosis in high peak VCD clonally derived cell lines (CDCLs). ER to Golgi vesicle mediated transport was found to be highly expressed in extended culture VCD CDCLs while networks involving endocytosis and oxidative stress response were significantly downregulated. Conclusion This investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production.

scholarly journals Cloning and characterization of small circular DNA from Chinese hamster ovary cells.

1984 ◽  
Vol 4 (1) ◽  
pp. 173-180 ◽  
Author(s):  
S W Stanfield ◽  
D R Helinski
Keyword(s):  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Single Copy ◽  
Chromosomal Dna ◽  
Cho Cell ◽  
Ovary Cells ◽  
Chromosomal Sequences

Small polydisperse circular (spc) DNA was isolated and cloned, using BglII from Chinese hamster ovary (CHO) cells. The properties of 47 clones containing at least 43 different BglII fragments are reported. The majority of the clones probably contain entire sequences from individual spcDNA molecules. Most of the clones were homologous to sequences in CHO cell chromosomal DNA, and many were also homologous to mouse LMTK- cell chromosomal sequences. The majority of homologous CHO cell chromosomal sequences were repetitive, although a few may be single copy. Only a small fraction of cloned spcDNA molecules were present in every cell; most occurred less frequently than once in 15 cells. Localization studies indicated that at least a portion of spcDNA is associated with the nucleus in CHO cells.

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