Article
Biochemical Research Methods
Yusuke Shibafuji, Nobuyoshi Nagao, Masafumi Yohda
Summary: Multi-omics analysis compared the effects of adding Tyr and Cystine in CHO cells. Adding Cystine resulted in decreased viability and productivity due to ER stress, ERAD, and apoptosis. On the other hand, adding Tyr suppressed ER stress and apoptosis by increasing ubiquinone biosynthesis. Simultaneously adding Tyr with Cystine improved growth, viability, and mAb productivity by activating GSH metabolism, suppressing ER stress and oxidative stress, reducing ERAD, and activating the tricarboxylic acid cycle.
BIOTECHNOLOGY JOURNAL
(2023)
Article
Biochemistry & Molecular Biology
Robert J. Edwards, Katayoun Mansouri, Victoria Stalls, Kartik Manne, Brian Watts, Rob Parks, Katarzyna Janowska, Sophie M. C. Gobeil, Megan Kopp, Dapeng Li, Xiaozhi Lu, Zekun Mu, Margaret Deyton, Thomas H. Oguin, Jordan Sprenz, Wilton Williams, Kevin Saunders, David Montefiori, Gregory D. Sempowski, Rory Henderson, Munir Alam, Barton F. Haynes, Priyamvada Acharya
Summary: The spike ectodomain of SARS-CoV-2 is destabilized by cold temperature storage but can be reversed by incubation at 37 degrees C or stabilizing its conformation in the 'down' state. This finding has significant implications for structural, functional, and vaccine studies related to the SARS-CoV-2 spike protein.
NATURE STRUCTURAL & MOLECULAR BIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Rita Agostinetto, Mara Rossi, Jessica Dawson, Angela Lim, Mirva H. Simoneau, Cyril Boucher, Bernhard Valldorf, Adin Ross-Gillespie, Joseph G. Jardine, Devin Sok, Dennis R. Burton, Thomas Hassell, Herve Broly, Wolf Palinsky, Philippe Dupraz, Mark Feinberg, Antu K. Dey
Summary: A novel transposon-based system was used to accelerate the generation of stable pools for monoclonal antibody production, providing clinical trial material within 4.5 months and meeting all pre-set product quality specifications.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Cell Biology
Yannic C. Bartsch, Xin Tong, Jaewon Kang, Maria Jose Avendano, Eileen F. Serrano, Tamara Garcia-Salum, Catalina Pardo-Roa, Arnoldo Riquelme, Yongfei Cai, Isabella Renzi, Guillaume Stewart-Jones, Bing Chen, Rafael A. Medina, Galit Alter
Summary: Despite evading neutralizing antibodies, the Omicron variant does not increase death rates in highly vaccinated populations, suggesting the presence of immune mechanisms beyond antibody-mediated virus neutralization. Vaccine-induced Spike protein-specific antibodies continue to drive Fc effector functions, indicating the potential of extraneutralizing antibodies in disease control.
SCIENCE TRANSLATIONAL MEDICINE
(2022)
Article
Biotechnology & Applied Microbiology
Nathaniel K. Hamaker, Lie Min, Kelvin H. Lee
Summary: The biomanufacturing industry is moving towards continuous processes, which may pose challenges due to fluctuations in host cell protein (HCP) levels. A study on Chinese hamster ovary (CHO) cells showed that HCP expression changes over time, but not as dramatically as originally expected.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
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
Virology
Liwei Li, Sina Qiao, Shumao Wang, Jiachen Liu, Kuan Zhao, Yanjun Zhou, Guoxin Li, Yifeng Jiang, Changlong Liu, Guangzhi Tong, Wu Tong, Fei Gao
Summary: This study successfully expressed ASFV p17 in CHO cells and generated a specific antibody against p17. The identified linear epitope of the antibody was conservative among different ASFV strains, indicating its potential for efficient detection and promising application prospects in future ASF vaccine research.
Article
Biotechnology & Applied Microbiology
Bassem Ben Yahia, Laetitia Malphettes, Elmar Heinzle
Summary: This study presents a systematic approach to predict CHO cell growth, metabolism, and mAb formation in biopharmaceutical production. It considers the impact of intracellular metabolites and successfully predicts the influence of new feeding strategies on cell growth and mAb production. The model developed shows promise for in silico prediction of optimal operating conditions.
METABOLIC ENGINEERING
(2021)
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
Medicine, Research & Experimental
Seo-Young Park, Susan Egan, Anthony J. Cura, Kathryn L. Aron, Xuankuo Xu, Mengyuan Zheng, Michael Borys, Sanchayita Ghose, Zhengjian Li, Kyongbum Lee
Summary: Reduction of monoclonal antibody disulfide bonds can impact therapeutic efficacy and safety, with potential factors including host cell proteome and stress response pathways. Analyzing Chinese hamster ovary cell protein abundances revealed distinctive characteristics in reduction-susceptible cell culture fluids, suggesting optimization of culture conditions may improve protein quality.
Article
Biotechnology & Applied Microbiology
Rajinder Kaur, Ritu Jain, Niharika Budholiya, Anurag S. Rathore
Summary: The study investigates the impact of long term culturing on cell morphology and altered cellular functions, which may lead to phenotypic drift and affect product yield and quality. The study highlights the use of cellular and analytical assay toolbox to define cell line stability and optimize culture performance and product quality.
BIOTECHNOLOGY LETTERS
(2023)
Article
Biotechnology & Applied Microbiology
Rin Miyajima, Hitomi Manaka, Tatsuya Honda, Noritaka Hashii, Masato Suzuki, Masahiro Komeno, Koichi Takao, Akiko Ishii-Watabe, Kazuei Igarashi, Toshihiko Toida, Kyohei Higashi
Summary: The depletion of polyamines in CHO cells increases IgG galactosylation and induces ER stress related to glycoprotein folding, leading to growth arrest. Introducing polyamines, especially SPD, to serum-free CHO culture medium may contribute to consistent antibody production and quality control.
JOURNAL OF BIOTECHNOLOGY
(2023)
Article
Chemistry, Analytical
Prity Kumari, Marianne Saldanha, Ratnesh Jain, Prajakta Dandekar
Summary: Addition of sugar osmolytes and formulation excipients can effectively reduce monoclonal antibody aggregation during cell culture, improving downstream processing and process yield.
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
(2023)
Article
Biotechnology & Applied Microbiology
Katharina Meyer, Ina G. Siller, Jana Schellenberg, Alina Gonzalez Salcedo, Doerte Solle, Jens Matuszczyk, Thomas Scheper, Janina Bahnemann
Summary: This study demonstrates the potential advantages of using the cold capture assay for monitoring cell culture productivity, showing that it is a time-effective and potentially cheaper alternative compared to traditional HPLC analysis. The results obtained from the cold capture assay were in great accordance with those from HPLC analysis throughout the cultivation process.
ENGINEERING IN LIFE SCIENCES
(2021)
Review
Biochemistry & Molecular Biology
Kusum Kharga, Lokender Kumar, Sanjay Kumar Singh Patel
Summary: Sepsis is a life-threatening condition characterized by an uncontrolled inflammatory response to an infectious agent and its antigens. Monoclonal antibody (mAb)-based therapeutic strategies are being explored as a treatment option for severe sepsis and septic shock.
Article
Biotechnology & Applied Microbiology
Stuart P. De Poi, Jianling Xie, C. Mark Smales, Christopher G. Proud
Summary: The study demonstrates that expressing certain constitutively active mutants of Rheb in CHO cells can lead to faster protein synthesis and increased secretion of valuable proteins, providing a simple approach to promoting cell growth and production of high commercial value secreted proteins in defined media.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Biotechnology & Applied Microbiology
Tanya J. Knight, Sarah Turner, Colin M. Jaques, Mark Smales
Summary: CHO cells are widely used for recombinant protein production, but some proteins are difficult to express in this system. This study found that proteasome activity in CHO cells is linked to their ability to produce recombinant proteins, and applying proteasome inhibitors during cell pool construction can enhance productivity of recombinant cell lines expressing difficult-to-express proteins.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
James D. Budge, Robert J. Young, Christopher Mark Smales
Summary: Transient gene expression in mammalian cells relies on efficient DNA delivery into the nucleus, which can be enhanced by NDPK-A protein with NLS. Co-expression of NDPK-A and EBNA-1 with oriP elements leads to improved transient recombinant protein yields despite attenuated cell growth. Combining these technologies results in superior protein yields compared to using either approach independently.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2021)
Review
Biotechnology & Applied Microbiology
Elizabeth Edwards, Maria Livanos, Anja Krueger, Anne Dell, Stuart M. Haslam, C. Mark Smales, Daniel G. Bracewell
Summary: Glycosylation plays a critical role in the quality and characteristics of therapeutic monoclonal antibodies in biologic manufacturing. Advances have been made in upstream processing, such as mammalian cell line engineering, to improve the predictability of glycosylation. Downstream processing strategies, including the use of novel technologies, are also being explored to control glycosylation. However, more research is needed to establish holistic and consistent manufacturing processes for antibody therapies.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Review
Biochemical Research Methods
Daniel G. Bracewell, Victoria Smith, Mike Delahaye, Mark C. Smales
Summary: The article discusses the development of HCP analysis and how insights from therapeutic monoclonal antibody preparations can inform the analysis of gene therapy viral vector products. While mass spectrometry approaches for HCP analysis in viral vector preparations have been established, they are yet to be widely applied in process or host cell line development. As these technologies are implemented and data becomes available, it will accelerate process development activities.
CURRENT OPINION IN BIOTECHNOLOGY
(2021)
Review
Biochemical Research Methods
Laura E. Dyball, C. Mark Smales
Summary: Exosomes are small spherical vesicles released into the extracellular environment via fusion of multivesicular bodies (MVBs) to the plasma membrane. They are involved in various physiological processes and have potential as biomarkers and therapeutic agents. However, there is still limited understanding of exosome biogenesis, manufacturing, characterization, and application methods.
BIOTECHNOLOGY JOURNAL
(2022)
Letter
Dermatology
John R. P. Knight, Christopher G. Proud, Giovanna Mallucci, Tobias von der Haar, C. Mark Smales, Anne E. Willis, Owen J. Sansom
JOURNAL OF INVESTIGATIVE DERMATOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Tanya J. Knight, Jane F. Povey, Davide Vito, Atul Mohindra, Colin M. Jaques, C. Mark Smales
Summary: In this study, the site of fragmentation of a model immunoglobulin G1 Fc-fusion protein was identified, and the observed clipping and aggregation were reduced by lowering the temperature and adding chemical reagents. The findings suggest that manipulating mRNA translation can improve protein folding and reduce the susceptibility to fragmentation.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Biotechnology & Applied Microbiology
Gary Montague, Olga Obrezanova, John Liddell, Alison Turner, Suzanne Aldington, Jamie Graham, Andy Racher, Susanne Joerg
Summary: This paper discusses a strategy for identifying decisions and bottlenecks in monoclonal antibody development and introduces a knowledge acquisition technique to enhance decision-making and understanding of the development supply chain.
FOOD AND BIOPRODUCTS PROCESSING
(2022)
Article
Biochemistry & Molecular Biology
Fabio Hedayioglu, Emma J. Mead, Patrick B. F. O'Connor, Matas Skiotys, Owen J. Sansom, Giovanna R. Mallucci, Anne E. Willis, Pavel Baranov, C. Mark Smales, Tobias von der Haar
Summary: The study validates two experimental tools for assessing ribosome binding to mRNAs, highlighting the usefulness of polysome profile reconstructions in evaluating dataset quality.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Biotechnology & Applied Microbiology
Lakshmeesha K. Nagappa, Wakana Sato, Farzana Alam, Kameshwari Chengan, Christopher M. Smales, Tobias von der Haar, Karen M. Polizzi, Katarzyna P. Adamala, Simon J. Moore
Summary: Cell-free gene expression systems are important tools in synthetic biology and industrial production. This study discovered a new, low-cost and widely available amino acid source that can replace commercial ones in these systems. The method is applicable to various cell-free gene expression systems.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Editorial Material
Biotechnology & Applied Microbiology
Anurag S. Rathore, C. Mark Smales, Andras Guttman
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Review
Biochemistry & Molecular Biology
Tobias von der Haar, Thomas E. Mulroney, Fabio Hedayioglu, Sathishkumar Kurusamy, Maria Rust, Kathryn S. Lilley, James E. Thaventhiran, Anne E. Willis, C. Mark Smales
Summary: In vitro transcribed, modified messenger RNAs (IVTmRNAs) have been widely used for COVID-19 vaccination and are being explored for other therapeutic applications. However, differences in genesis pathways, cell entry routes, and modifications affect how IVTmRNAs interact with cellular machinery and how efficiently they are translated. Understanding the similarities and differences in translation between IVTmRNAs and native mRNAs is crucial for improving their therapeutic efficacy.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2023)
Article
Biotechnology & Applied Microbiology
James D. Budge, Joanne Roobol, Gurdeep Singh, Theo Mozzanino, Tanya J. Knight, Jane Povey, Andrew Dean, Sarah J. Turner, Colin M. Jaques, Robert J. Young, Andrew J. Racher, C. Mark Smales
Summary: CHO cells are commonly used to produce complex recombinant biotherapeutics like monoclonal antibodies. A selection marker system based on the metabolic needs of CHO cells for proline production has been developed to enhance recombinant protein productivity through engineering lipid metabolism.
METABOLIC ENGINEERING COMMUNICATIONS
(2021)