Review
Biochemistry & Molecular Biology
Katja Glinsek, Kristof Bozovicar, Tomaz Bratkovic
Summary: CRISPR technology has made significant progress in CHO cell line engineering, improving protein production and product quality. It allows for glycosylation modulation, productivity enhancement, tackling adventitious agents, elimination of problematic host cell proteins, development of antibiotic-free selection systems, site-specific transgene integration, and CRISPR-mediated gene activation and repression. The review highlights the potential of CRISPR technology in CHO cell line genome editing and epigenetic engineering for more efficient and cost-effective development of biopharmaceuticals, ensuring the safety and quality of the final product.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Review
Biotechnology & Applied Microbiology
Jun-He Zhang, Lin-Lin Shan, Fan Liang, Chen-Yang Du, Jing-Jing Li
Summary: Recombinant antibodies are a rapidly developing class of therapeutic agents, but high-level expression and production of these antibodies remain a major challenge. This review provides a comprehensive overview of various strategies for optimizing gene sequences, expression vectors, host cell lines, and glycosylation modifications to improve the efficiency and quality of recombinant antibody production.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Hyeonjin Cha, Ju-Hyun Park
Summary: The addition of 30Kc19 alpha protein was found to enhance the production of EPO in CHO cell culture, with potential advantages in downstream purification process. 30Kc19 alpha could be mass-produced as a recombinant protein and delivered intracellularly to improve EPO production through metabolic pathways. The results suggest that 30Kc19 alpha has promising potential for manufacturing biopharmaceutical proteins.
APPLIED SCIENCES-BASEL
(2021)
Article
Biochemical Research Methods
Feiyang Zheng, Yoshinori Kawabe, Mai Murakami, Mamika Takahashi, Kyoka Nishihata, Souichiro Yoshida, Akira Ito, Masamichi Kamihira
Summary: The study developed a retrotransposon-based vector system that achieved integration of multiple transgenes into host genome and successfully introduced scFv-Fc into Chinese hamster ovary cells. Separately expressing ORF1 and ORF2 of LINE-1 efficiently regulated retrotransposition and improved specific scFv-Fc productivity.
BIOTECHNOLOGY JOURNAL
(2021)
Article
Biotechnology & Applied Microbiology
Honggi Min, Seul Mi Kim, Dongwoo Kim, Solhwi Lee, Sumin Lee, Jae Seong Lee
Summary: In this study, a hybrid cell line development platform was developed to increase the productivity of targeted integrants through the amplification of transgene copies. The results showed that targeted integrants exhibited a 3.6-fold increase in EGFP expression and increased copy numbers of DHFR and EGFP when exposed to 200 nM MTX. A single-step MTX amplification increased the specific monoclonal antibody productivity by 2.8-fold. The study provides a new strategy for increasing the productivity of CHO cell lines.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Review
Biotechnology & Applied Microbiology
Laura Bryan, Martin Clynes, Paula Meleady
Summary: CHO cells are commonly used host cell lines for producing human therapeutic proteins. Research has focused on improving growth, titre, and productivity of CHO cells to reduce production costs. Recent interest in CHO cell engineering has led to studies on post-translational modifications affecting protein functionality.
BIOTECHNOLOGY ADVANCES
(2021)
Article
Biotechnology & Applied Microbiology
Thu M. Cao, Dayue Chen, Gavin C. Barnard, Amy Shen
Summary: This study explores the potential of using CHO cells for rAAV production and identifies cap protein expression as a limiting factor. By increasing cap protein expression, rAAV titers in CHO cells were comparable to those in HEK-derived rAAV, providing a foundation for potential rAAV production in CHO cells.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Review
Biotechnology & Applied Microbiology
Weifeng Li, Zhenlin Fan, Yan Lin, Tian-Yun Wang
Summary: This review highlights the importance of serum-free medium (SFM) in the production of recombinant therapeutic proteins (RTPs) using Chinese hamster ovary (CHO) cells, with nearly 70% of RTPs currently produced in this manner. It discusses the history, key components, preparation, and experimental design of SFM, as well as the potential benefits of small molecule compound additives in improving RTP yield and quality. Lastly, it explores the future prospects of using SFM with CHO cells for RTP production.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2021)
Article
Biochemical Research Methods
Jiang-Tao Lu, Meng-Ke Xiao, Ying-Ying Feng, Xiao-Yin Wang, Le-Le Qiu, Yu-Rong Chai, Tian-Yun Wang, Yan-Long Jia
Summary: The small-molecule compound apilimod was found to positively influence recombinant protein expression in CHO cells by selectively blocking the cell cycle and regulating key regulators involved in cell proliferation and autophagy, suggesting its potential as a novel enhancer in CHO cell engineering.
BIOTECHNOLOGY JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Jesus E. Martinez-Lopez, Orla Coleman, Paula Meleady, Martin Clynes
Summary: MicroRNAs play an increasingly important role in enhancing relevant pathways during CHO cell line development and optimizing biopharmaceutical production processes. MiR-31* has been shown to boost oxidative phosphorylation in mitochondria, leading to increased specific productivity in CHO cells.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Lara Bezjak, Vivian Erklavec Zajec, Spela Baebler, Tjasa Stare, Kristina Gruden, Andrej Pohar, Uros Novak, Blaz Likozar
Summary: This work improved the kinetic model based on metabolic and glycan reaction networks of CHO cells by integrating transcriptomic data, resulting in better predictions of metabolic concentrations in the Fed-Batch process, marking a significant advancement in modeling techniques.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Biotechnology & Applied Microbiology
Ioscani Jimenez del Val, Sarantos Kyriakopoulos, Simone Albrecht, Henning Stockmann, Pauline M. M. Rudd, Karen M. M. Polizzi, Cleo Kontoravdi
Summary: Metabolic modeling is important for characterizing biopharmaceutical cell culture processes. However, current metabolic models of CHO cells face challenges in terms of size and interpretation. To address these challenges, the CHOmpact model was developed as a reduced metabolic network with improved interpretability and physiological consistency. This model provides a platform for developing dynamic metabolic models to optimize biopharmaceutical cell culture processes.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Biochemistry & Molecular Biology
Grace Yao, Kathryn Aron, Michael Borys, Zhengjian Li, Girish Pendse, Kyongbum Lee
Summary: The research identified potential high productivity indicators in CHO cell clones, such as citrate, which improved qP by up to 490% when used as a medium additive. This demonstrates the potential of metabolomics in discovering novel metabolite additives for higher volumetric productivity in biologics production processes.
Article
Biotechnology & Applied Microbiology
Benjamin Strain, James Morrissey, Athanasios Antonakoudis, Cleo Kontoravdi
Summary: This study evaluates the reliability of Chinese hamster ovary (CHO) cell metabolic models and introduces a new model that outperforms the existing ones in predicting extracellular phenotypes and intracellular fluxes. However, there is still room for improvement in predicting intracellular metabolic states. This work provides an updated CHO cell model and lays the foundation for further research in flux analysis techniques.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Song-Min Schinn, Carly Morrison, Wei Wei, Lin Zhang, Nathan E. Lewis
Summary: Genome-scale metabolic models can accurately predict individual amino acid concentrations in culture medium throughout the production process by integrating machine learning, enabling real-time control of nutrient feeding.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)