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
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
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)
Article
Biotechnology & Applied Microbiology
Eleftheria Skrika-Alexopoulos, C. Mark Smales
Summary: Exosomes were isolated from CHO cells using a novel method and characterized to determine their protein and RNA content. These findings contribute to the future research and improvement of exosome manufacturing from CHO cells.
BIOTECHNOLOGY LETTERS
(2023)
Review
Biochemical Research Methods
James Donaldson, Dirk-Jan Kleinjan, Susan Rosser
Summary: Fed-batch culture of Chinese hamster ovary (CHO) cells is the most commonly used method for producing biopharmaceuticals. Static CHO cell-line engineering has improved productivity and quality, but conditions change during fed-batch culture. Dynamic engineering strategies can optimize traits by implementing synthetic sense-and-respond programs.
CURRENT OPINION IN BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Kexue Liang, Hongzhen Luo, Qi Li
Summary: In this study, the effects of different perfusion culture strategies on process stability, product titer, and product quality were investigated. Continuous perfusion was found to be more effective in maintaining high levels of cell density, product titer, and quality compared to intermittent perfusion culture. Furthermore, the study optimized the perfusion culture conditions by adjusting perfusion rates and culture temperature. The optimized strategy resulted in significantly higher product titer and maintained excellent product quality.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
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)
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)
Article
Biotechnology & Applied Microbiology
Pavlos Kotidis, Roberto Donini, Johnny Arnsdorf, Anders Holmgaard Hansen, Bjorn Gunnar Rude Voldborg, Austin W. T. Chiang, Stuart M. Haslam, Michael Betenbaugh, Ioscani Jimenez del Val, Nathan E. Lewis, Frederick Krambeck, Cleo Kontoravdi
Summary: Researchers constructed a comprehensive glycosylation reaction network, CHOGlycoNET, based on 200 glycan datasets from CHO cell lines. They used machine learning and dimensionality reduction techniques to explore the distribution of mannosidase and glycosyltransferase enzymes in the Golgi apparatus and identify key reactions. CHOGlycoNET can accelerate glycomodel development and predict the effect of glycoengineering strategies.
METABOLIC ENGINEERING
(2023)
Review
Biotechnology & Applied Microbiology
Tingting Xu, Jihong Zhang, Tianyun Wang, Xiaoyin Wang
Summary: This review focuses on the aggregation of recombinant therapeutic antibodies (RTAs), discussing factors causing aggregation during RTA production and strategies for overcoming RTA aggregation.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Nanoscience & Nanotechnology
Filip Stolinski, Anna Rybinska-Fryca, Maciej Gromelski, Alicja Mikolajczyk, Tomasz Puzyn
Summary: The purpose of the research is to develop a user-friendly web-based application that can predict the cytotoxicity of nanomaterials. The application is not only suitable for chemoinformatics specialists but also for experts in nanotechnology or toxicology.
Article
Biotechnology & Applied Microbiology
Dongil Kim, Su Hyun Kim, Chansik Yoon, Gyun Min Lee
Summary: The use of CRISPR/Cas9-based single-gene knockout cell library was able to identify KO target genes conferring resistance to iHDACs, leading to improved therapeutic protein production.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Dongil Kim, Chansik Yoon, Gyun Min Lee
Summary: In this study, the benzamide-based histone deacetylase inhibitor CI994 was found to increase monoclonal antibody production in Chinese hamster ovary cells by remodeling chromatin and enhancing gene transcription.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Biochemical Research Methods
Patrick G. Schweickert, Ning Wang, Stephanie L. Sandefur, Michael E. Lloyd, Stephen F. Konieczny, Christopher. C. Frye, Zhuo Cheng
Summary: The study demonstrates the successful application of CRISPR/Cas12a for generating CHO KO cell lines with improved product quality, with efficiency dependent on the targeting RNA used; small screens of clonally derived cell lines can be used to generate CHO KO cell lines, and a novel bulk culture analysis approach is proposed for assessing CRISPR RNA efficiency and determining ideal screen sizes; Cas12a can be directly integrated into the cell line generation process with no negative impact on titer or screen size.
BIOTECHNOLOGY JOURNAL
(2021)
Article
Biotechnology & Applied Microbiology
Shivani Gupta, Bhavana Shah, Coral Shek Fung, Pik Kay Chan, Devin L. Wakefield, Scott Kuhns, Chetan T. Goudar, James M. Piret
Summary: Since 2015, more than 34 biosimilars have been approved by the FDA, leading to a renewed focus on technology development for therapeutic protein production. The genetic differences between host cell lines used in biologics manufacturing pose a challenge. To address this, a CHO cell line was engineered to produce monoclonal antibodies with murine glycans, aiming to match the reference product expressed in murine cells. This study demonstrates the feasibility of using CHO cells to produce mAbs with murine glycans, potentially reducing uncertainty in biosimilarity development.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Ioanna Tzani, Craig Monger, Krishna Motheramgari, Clair Gallagher, Ryan Hagan, Paul Kelly, Alan Costello, Justine Meiller, Patrick Floris, Lin Zhang, Martin Clynes, Jonathan Bones, Niall Barron, Colin Clarke
BIOTECHNOLOGY AND BIOENGINEERING
(2020)
Article
Biotechnology & Applied Microbiology
Krishna Motheramgari, Ricardo Valdes-Bango Curell, Ioanna Tzani, Clair Gallagher, Marina Castro-Rivadeneyra, Lin Zhang, Niall Barron, Colin Clarke
BIOTECHNOLOGY AND BIOENGINEERING
(2020)
Article
Biotechnology & Applied Microbiology
Ioanna Tzani, Nicolas Herrmann, Sara Carillo, Cathy A. Spargo, Ryan Hagan, Niall Barron, Jonathan Bones, W. Shannon Dillmore, Colin Clarke
Summary: The study used single-cell RNA sequencing to investigate the production instability of a clonally derived CHO cell line, revealing that heavy chain gene expression was significantly lower compared to that of the light chain. The evolution of the cell line showed a reduction in heavy and light chain gene expression, while genes involved in oxidative stress and apoptosis response increased in expression as cells progressed along the trajectory. Future studies utilizing this technology have the potential to enhance understanding of efficient manufacturing performance and product quality in CHO cell lines.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Biotechnology & Applied Microbiology
Laura Bryan, Michael Henry, Niall Barron, Clair Gallagher, Ronan M. Kelly, Christopher C. Frye, Matthew D. Osborne, Martin Clynes, Paula Meleady
Summary: The study identified miR-200a as a potential regulator of the unfolded protein response (UPR) in CHO cells, affecting protein folding and impacting Qp phenotype, suggesting it as a potential target for engineering industrially attractive CHO cell phenotypes.
BIOTECHNOLOGY LETTERS
(2021)
Article
Biochemistry & Molecular Biology
Lisa Strasser, Stefano Boi, Felipe Guapo, Nicholas Donohue, Niall Barron, Alana Rainbow-Fletcher, Jonathan Bones
Summary: AAV vectors are crucial for gene therapy, but manufacturing remains a challenge. This study investigated the host cell proteome of AAV5-producing cells, uncovering differential expression of proteins related to cellular metabolism, proliferation, cell death, as well as endocytosis and lysosomal degradation.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Medicine, Research & Experimental
Filippos T. Charitidis, Elham Adabi, Frederic B. Thalheimer, Colin Clarke, Christian J. Buchholz
Summary: Quantifying gene expression in individual cells can improve our understanding of complex genetically engineered cell products like CAR T cells. By using single-cell RNA sequencing, researchers were able to monitor the delivery of the CD19-CAR gene and identify a trimodal expression of CAR and CD8A within cells, leading to a reliable distinction between transduced and untransduced cells during CAR T cell generation. The study also revealed a high selectivity of on-target cells and upregulation of restriction factors in CAR-negative cells.
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
(2021)
Article
Virology
Marco Silvano, Ricardo Correia, Nikolaus Virgolini, Colin Clarke, Paula M. Alves, Ines A. Isidro, Antonio Roldao
Summary: This study investigates the gene expression patterns associated with high-producer insect cells adapted to neutral pH during the production of influenza HA-VLPs. The results show differential expression of host cell genes and baculovirus genes, with enrichment of pathways related to metabolism and protein folding. These findings contribute to our understanding of the biological mechanisms of insect cells during baculovirus-mediated expression.
Article
Biochemical Research Methods
Nga Lao, Niall Barron
Summary: m6A modification on eukaryotic mRNA plays a crucial role in cellular processes. This study demonstrates that manipulating the expression of m6A readers, YTHDF1, 2, and 3, can enhance recombinant protein yield and viral protein expression in mammalian cells. These findings suggest the potential of utilizing epitranscriptomic-based approaches for improving protein production and viral vector yield in host cell line engineering.
BIOTECHNOLOGY JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Nikolaus Virgolini, Marco Silvano, Ryan Hagan, Ricardo Correia, Paula M. Alves, Colin Clarke, Antonio Roldao, Ines A. Isidro
Summary: Using single-cell RNA sequencing, this study performed transcriptomics analysis of Sf9 insect cells during the production of rAAV2. The study found an increase in transcriptional heterogeneity during infection, which was linked to the expression of baculovirus genes and rAAV2 transgenes. Additionally, the study highlighted limitations of the dual-baculovirus system in producing rAAV2 particles.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)