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)
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
Simon Fischer, Sven Mathias, Anna Stadermann, Shumin Yang, Valerie Schmieder, Nikolas Zeh, Nicoletta Schmidt, Patrick Richter, Sara Wright, Eike Zimmermann, Yan Ley, Julia Meer, Thomas Hartsch, Christian Bernloehr, Kerstin Otte, Harald Bradl, Martin Gamer, Patrick Schulz
Summary: A study identified a recombinant CHO cell line with unusually high levels of NGNA sialylation, and the root cause was found through comprehensive multi-OMICs experimental analyses, involving the spontaneous expression of the CMAH gene, microRNA miR-111 regulation, and a point mutation in the SDK1 gene.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
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)
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)
Review
Biotechnology & Applied Microbiology
Melanie Nguyen, Aline Zimmer
Summary: Chinese hamster ovary (CHO) cells are widely used in the biopharmaceutical industry for producing recombinant proteins. The proteomic studies on CHO cells have provided valuable insights into the protein clusters, regulatory networks, and pathways governing phenotypic changes, which can be utilized to optimize CHO-based bioprocesses.
BIOTECHNOLOGY ADVANCES
(2023)
Review
Biotechnology & Applied Microbiology
Nicolas Marx, Peter Eisenhut, Marcus Weinguny, Gerald Klanert, Nicole Borth
Summary: Recent advances have greatly improved our understanding of Chinese hamster ovary (CHO) cells as hosts for producing biopharmaceuticals. Despite progress in characterizing CHO cells on different omics levels, our understanding of its epigenetic regulation is still incomplete. This review provides an overview of epigenetic regulation in CHO cells and discusses its significance in shaping cell phenotype. The review also presents current state-of-the-art technologies for manipulating epigenetic networks and controlling gene expression.
BIOTECHNOLOGY ADVANCES
(2022)
Review
Biotechnology & Applied Microbiology
Wenwen Yang, Junhe Zhang, Yunxi Xiao, Wenqing Li, Tianyun Wang
Summary: Chinese hamster ovary (CHO) cells are commonly used for recombinant protein expression in the pharmaceutical industry. Cell line development (CLD) is a critical step in producing therapeutic proteins, but the screening process is time-consuming and determines the level of high-yield cell lines. This review provides an overview of preliminary screening methods for high-yield cell pools, as well as high-throughput methods for isolating high-yield cell clones and increasing productivity and stability.
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
Multidisciplinary Sciences
Ngan T. B. Nguyen, Jianer Lin, Shi Jie Tay, Mariati, Jessna Yeo, Terry Nguyen-Khuong, Yuansheng Yang
Summary: In this study, the researchers manipulated the N-glycan structures on therapeutic antibodies by overexpressing various glycosyltransferase genes in CHO cells, resulting in antibodies with different N-linked glycan structures. This approach opens up new possibilities for developing novel recombinant antibodies for addressing specific therapeutic applications.
SCIENTIFIC REPORTS
(2021)
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
Biochemical Research Methods
Aleksandra Stec, Monika Targonska, Edyta Karkosinska, Monika Slowik, Agata Ploska, Leszek Kalinowski, Bartosz Wielgomas, Krzysztof Waleron, Jacek Jasiecki, Szymon Dziomba
Summary: The overproduction of proteins in Chinese hamster ovary (CHO) cells can induce the secretion of extracellular vesicles (EVs), potentially leading to the packaging of overexpressed proteins into exosomes. These findings are crucial for the manufacturing of therapeutic drugs in CHO expression systems, as there is a risk of product loss during downstream processing of culture medium and the use of exosomes as nanocarriers for therapeutic proteins.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2023)
Article
Biotechnology & Applied Microbiology
Hye-Jin Han, Jin-Hyuk Lim, Hyun-Myoung Cha, Ji-Hun Lee, Dong-Il Kim
Summary: Sialylation, a critical quality attribute in the pharmacokinetics of recombinant proteins, is enhanced by glucocorticoids (GCs) which increase sialylation levels and total sialic acid contents, thereby improving the quality of biopharmaceuticals.
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
(2021)
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)
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)