Conversion of a CHO cell culture process from perfusion to fed-batch technology without altering product quality

Meuwly, F.; Weber, Urs; Ziegler, Thomas R.; Gervais, Annick; Mastrangeli, Renato; Crisci, C.; Rossi, Mara; Bernard, A.; Stockar, Urs von; Kadouri, A.

doi:10.1016/j.jbiotec.2005.10.013

Cited by 46 publications

(36 citation statements)

References 8 publications

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“…For a new molecule without prior history in development, there is more freedom in process selection, since acceptable ranges of product quality attributes will be based on what is produced from the process chosen. However, when changing processes in late stage process development, extensive studies will have to be done to ensure comparability [46,47]. In those cases, the difference observed between different operational modes will have to be carefully evaluated, and extended characterization is typically required to understand the impact of them.…”

Section: Product Quality Attributes Comparison Between Different Opermentioning

confidence: 99%

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Hoshan

Chen

2016

Bioprocess Biosyst Eng

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In this study, we discussed the development and optimization of an intensified CHO culture process, highlighting medium and control strategies to improve lactate metabolism. A few strategies, including supplementing glucose with other sugars (fructose, maltose, and galactose), controlling glucose level at <0.2 mM, and supplementing medium with copper sulfate, were found to be effective in reducing lactate accumulation. Among them, copper sulfate supplementation was found to be critical for process optimization when glucose was in excess. When copper sulfate was supplemented in the new process, two-fold increase in cell density (66.5 ± 8.4 × 10(6) cells/mL) and titer (11.9 ± 0.6 g/L) was achieved. Productivity and product quality attributes differences between batch, fed-batch, and concentrated fed-batch cultures were discussed. The importance of process and cell metabolism understanding when adapting the existing process to a new operational mode was demonstrated in the study.

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Section: Product Quality Attributes Comparison Between Different Opermentioning

confidence: 99%

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Hoshan

Chen

2016

Bioprocess Biosyst Eng

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“…For many years the biopharmaceutical industry was aiming to increase specific productivity to minimize production cost-of-goods while maintaining product quality 30 . Several parameters including the biology of the production cell line, product quality and stability, manufacturing capacity, process scalability, volumetric productivity, and unit cost determine the selection of the production process, namely fed-batch or perfusion 31 . Due to product stability issues, many recombinant proteins, such as insulin and interferons, were produced in perfusion in the early stages of the biotechnology industry 32 .…”

Section: Cell Culture Process Optimizationmentioning

confidence: 99%

“…Despite the substantially increased productivity of fed-batch processes and manufacturing capacity, the latter has become an issue because of the constantly increasing demand of specific and thus efficacious biotherapeutics 9,31 . Further capacity increases by either building new facilities or by process intensification (eg.…”

Section: Cell Culture Process Optimizationmentioning

confidence: 99%

“…In addition to the host cell genome, the cell culture process conditions assuring cell growth and protein expression shape the protein quality attributes as well. According to the common phrase 'the process is the product', the metabolism of the cells closely depends on the culture conditions, including the pH 17 , the temperature 49 , the oxygen tension 17 , the CO 2 content in the culture broth 50 , as well as the type of process, namely perfusion or fed-batch mode 31 . Different metabolic states, which result from differences in culture parameters, very likely express proteins with altered quality attributes.…”

Section: Cell Culture Process Optimizationmentioning

confidence: 99%

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Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

Biotechnology Progress

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Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

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“…In addition, perfusion culture is beneficial for the purity and quality of the product, especially those bio-products that are sensitive to protease and glycosidase (Rastilho et al 2002;Ryll et al 2000). However, the perfusion process has a limited scale-up capability when compared with the fed-batch process, which has scale-up potential to typical bioreactor sizes of 20 m 3 or more (Meuwly et al 2006;John et al 2006). In this work, we evaluated batch, fed-batch, and perfusion operations to optimize the process for rapid production of anti-EGFRvIII C12 antibody from recombinant CHO cells cultivated in serumfree medium for pre-clinical and early stage clinical trials.…”

Section: Introductionmentioning

confidence: 99%

Bioprocess development for the production of mouse-human chimeric anti-epidermal growth factor receptor vIII antibody C12 by suspension culture of recombinant Chinese hamster ovary cells

Deng

Wang

et al. 2011

Cytotechnology

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The mouse-human chimeric anti-epidermal growth factor receptor vIII (EGFRvIII) antibody C12 is a promising candidate for the diagnosis of hepatocellular carcinoma (HCC). In this study, 3 processes were successfully developed to produce C12 by cultivation of recombinant Chinese hamster ovary (CHO-DG44) cells in serum-free medium. The effect of inoculum density was evaluated in batch cultures of shaker flasks to obtain the optimal inoculum density of 5 9 10 5 cells/mL. Then, the basic metabolic characteristics of CHO-C12 cells were studied in stirred bioreactor batch cultures. The results showed that the limiting concentrations of glucose and glutamine were 6 and 1 mM, respectively. The culture process consumed significant amounts of aspartate, glutamate, asparagine, serine, isoleucine, leucine, and lysine. Aspartate, glutamate, asparagine, and serine were particularly exhausted in the early growth stage, thus limiting cell growth and antibody synthesis. Based on these findings, fedbatch and perfusion processes in the bioreactor were successfully developed with a balanced amino acid feed strategy. Fed-batch and especially perfusion culture effectively maintained high cell viability to prolong the culture process. Furthermore, perfusion cultures maximized the efficiency of nutrient utilization; the mean yield coefficient of antibody to consumed glucose was 44.72 mg/g and the mean yield coefficient of glutamine to antibody was 721.40 mg/g. Finally, in small-scale bioreactor culture, the highest total amount of C12 antibody (1,854 mg) was realized in perfusion cultures. Therefore, perfusion culture appears to be the optimal process for small-scale production of C12 antibody by rCHO-C12 cells.

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Conversion of a CHO cell culture process from perfusion to fed-batch technology without altering product quality

Cited by 46 publications

References 8 publications

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Tailoring recombinant protein quality by rational media design

Bioprocess development for the production of mouse-human chimeric anti-epidermal growth factor receptor vIII antibody C12 by suspension culture of recombinant Chinese hamster ovary cells

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