Enhancing and stabilizing monoclonal antibody production by Chinese hamster ovary (CHO) cells with optimized perfusion culture strategies

Abstract

The perfusion medium is critical in maintaining high cell concentration in cultures for the production of monoclonal antibody by Chinese hamster ovary cells. In this study, the effects of perfusion culture strategies when using different media on the process stability, product titer, and product quality were investigated in 3-L bioreactor. The results indicated that continuous perfusion could maintain higher levels of cell density, product titer, and quality in comparison with those of the intermittent perfusion culture. Next, the perfusion culture conditions with different perfusion rates and temperature reduction methods were further optimized. When combining the high perfusion rates and delayed reduction of culture temperature at day 6, the product titer reached a higher level of 16.19 g/L with the monomer relative abundant of 97.6%. In this case, the main peak of the product reached 56.3% and the total N-glycans ratio was 95.2%. To verify the effectiveness of the optimized perfusion culture in a larger scale, a 200-L bioreactor was used to perform and the final product titer reached the highest level of 16.79 g/L at day 16. Meanwhile, the product quality (monomer abundant of 97.6%, main peak of 56.3%, and N-glycans ratio of 96.5%) could also be well maintained. This study provided some guidance for the high-efficient production of monoclonal antibody by CHO cells via optimized perfusion culture strategy.

Effect of plant produced Anti-hIL-6 receptor antibody blockade on pSTAT3 expression in human peripheral blood mononuclear cells

“…Other plant produced proteins expressed showed yields of 18.49 mg/kg of human interleukin-6 58 , 3 mg/kg of cocaine-hydrolase Fc protein 59 , 730 mg/kg of CMG2-Fc protein 60 . These short time periods provide essential advantages of protein expression in terms of speed over other expression approaches including mammalian cells 61 – 64 and stable plant expression 65 – 67 systems. The significant reduction in antibody expression after 5 dpi might be due to the progressive development of necrosis in the infiltrated leaves.…”

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

“…Perfusion (PER) culture is a continuous culture mode, in which fresh medium is continuous to be added into the bioreactor and the product is continuously harvested. Therefore, PER culture is a relatively new technique to maintain cell viability and harvest high-quality proteins (Liang et al 2023 ; Schwarz et al 2023 ). PER culture also provides a narrower residence time distribution for products, especially unstable RTPs like FVIII, limiting exposure to extracellular enzymes (e.g., proteases, glycosidases, sialidases) as well as the extracellular environment (pH, temperature) (Coronel et al 2020 ).…”

“…With the progress and development of medium optimization and cell culture technology, the expression levels of RTPs in CHO cells (Gbico, #A13696-01) were significantly increased and some monoclonal antibody titers exceeded 10 g/L under the conditions of fed-batch culture and perfusion culture (Handlogten et al 2018 ; Srila et al 2023 ; Xu et al 2023 ). By 2020, more than 70 monoclonal antibodies (mAbs) are already on the market and global sales will reach $125 billion (Liang et al 2023 ; Urquhart 2021 ). Biopharmaceutical market approval from 2014 to 2018 showed that more than 80% of newly approved biopharmaceuticals were produced with CHO cells, and that number had risen to 89% until 2022 (Walsh and Walsh 2022 ).…”

Enhancing recombinant antibody yield in Chinese hamster ovary cells

“…To validate the effectiveness of the optimized perfusion culture on a larger scale, a 200-L bioreactor was utilized, resulting in the highest product titer reaching 16.79 g/L by day 16. This study offers valuable insights for achieving highly efficient production of mAbs by CHO cells through an optimized perfusion culture strategy [ 78 ].…”