Article
Biochemical Research Methods
Elizabeth Greene, Daniela Cazacu, Ninkka Tamot, Steven Castellano, Akshita Datar, Anthony Kronkaitis, Douglas Gebhard, Jon Reed, Paul Mawson, Lore Florin, Nicholas Rossi, Anthony Lauer, Laura Juckem, Andrew Nixon, Till Wenger, Saurabh Sen
Summary: This study presents the optimization of parameters for TGE for antibodies from CHO cells, resulting in a uniquely optimized process with higher titer and reduced costs and time. It also investigates the amount of coding DNA used in TGE and the influence of kinetics and size of the transfection complex on the in vitro efficiency of the transfection, presenting the first report of an optimized TGE platform using Filler DNA for the screening and production of therapeutic mAbs in an early drug discovery setting.
BIOTECHNOLOGY JOURNAL
(2021)
Article
Pharmacology & Pharmacy
Ly Porosk, Jekaterina Nebogatova, Heleri Heike Hark, Birgit Vunk, Piret Arukuusk, Urve Toots, Mart Ustav, Ulo Langel, Kaido Kurrikoff
Summary: Therapeutic proteins are at the forefront of innovation in pharmaceutical medicine, but their industrial production poses technical challenges. This study aimed to find a fast, microliter-scale transfection assay for predicting protein expression in transient production settings. The results showed that commonly used microliter-scale transfection efficacy assays fail to predict performance in large-scale protein production. Additionally, cell-penetrating peptide-based transfection achieved significantly higher protein yields compared to other methods.
Article
Biochemistry & Molecular Biology
Ziva Svab, Luca Braga, Corrado Guarnaccia, Ivan Labik, Jeremias Herzog, Marco Baralle, Mauro Giacca, Natasa Skoko
Summary: In this study, several human miRNAs were identified to increase specific and overall production of Erythropoietin and Etanercept in CHO cells. Overexpression of miR-574-3p was found to improve productivity by up to 2-fold, with p300 identified as a main target. The findings highlight the importance of genomic sequences in pri-miRNA processing, offering a cost-effective strategy for efficient miRNA engineering in CHO cells.
Article
Biotechnology & Applied Microbiology
Noriko Yamano-Adachi, Yuto Nakanishi, Wataru Tanaka, YuanShan Lai, Masahiro Yamazaki, Laura Zenner, Hirofumi Hata, Takeshi Omasa
Summary: This study investigated the effects of artificial changes in the chromosome number on productivity in CHO cells. It was found that cell populations with a diverse chromosome number distribution exhibited higher antibody productivities. Additionally, inducing aneuploid cells using 3-aminobenzamide increased antibody productivity. Furthermore, using host cells with an average chromosome number of 37 significantly improved antibody productivity and increased the integration of exogenous genes into the genome.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Chemistry, Multidisciplinary
Shanhe Li, Ze Wen, Behafarid Ghalandari, Tianhao Zhou, Antony R. Warden, Ting Zhang, Peng Dai, Youyi Yu, Wenke Guo, Mofang Liu, Haiyang Xie, Xianting Ding
Summary: The single-cell transfection analysis chip (scTAC) is a new technology based on the in situ microchip immunoblotting method, which can rapidly and accurately analyze exogenous gene expression in individual host cells, and reveal the expression level relationship between reporter genes and target genes.
ADVANCED MATERIALS
(2021)
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
Biotechnology & Applied Microbiology
Qinghao Zhang, Bo Jiang, Luke Nelson, Steven Huhn, Zhimei Du, Lawrence A. Chasin
Summary: CHO8A, a multiauxotrophic mutant of CHO-K1 cells, was created by restoring the prototrophy of the cells through transfection of complementary DNA-based genes. This cell line allows for the selection of transfectant clones with multiple polypeptide genes, as well as the single-step selection of high producers of a specific protein.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Biochemistry & Molecular Biology
Jaemun Kim, Ji Yul Kim, Hyeonkyeong Kim, Eunsil Kim, Soonyong Park, Kyoung-Hwa Ryu, Eun Gyo Lee
Summary: Transient gene expression using DC-Chol/DOPE cationic liposomes and pDNA in Chinese hamster ovary suspension cells was established in this study. By optimizing complexation solutions, high levels of human Fc protein and immunoglobulin G were successfully produced. This acidic and alkaline lipoplex formation could potentially be an effective gene delivery strategy for biopharmaceutical production.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biochemistry & Molecular Biology
Raffaele Altamura, Jiten Doshi, Yaakov Benenson
Summary: Cell line development is crucial in biopharmaceutical manufacturing, but current protocols rely on random transgene integration. A novel strategy has been proposed to reduce screening effort, improve stability, and predict product titers through engineered multi-copy gene constructs.
NUCLEIC ACIDS RESEARCH
(2022)
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
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
Biochemistry & Molecular Biology
Shaowei Sui, Hao Wang, Jiajie Song, Wanyi Tai
Summary: Transient expression is the only way to quickly obtain a small scale of antibodies for biomedical research and therapeutic evaluation. A novel spermine-based lipid, sperminoliposome, was developed as an effective and cheap transfection agent for antibody expression. Compared to PEI, sperminoliposome achieved a 3 times higher yield of antibody during transient expression in suspension cells, confirming its feasibility and efficiency for rapid antibody production.
BIOORGANIC & MEDICINAL CHEMISTRY
(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
Tobias Jerabek, Florian Klingler, Nadja Raab, Nikolas Zeh, Jens Pfannstiel, Kerstin Otte
Summary: The production of biopharmaceuticals relies on robust cell systems, such as Chinese hamster ovary cells (CHO), that can produce proteins at high levels and survive in stressful environments. The study conducted pathway enrichment analysis of the CHO surfaceome and observed enrichment of growth/survival pathways, providing novel insights into how cell surface receptors and ligand-mediated signaling enable cells to grow and survive in a bioprocess environment. This research can serve as a foundation for future studies and optimization strategies to enhance cellular growth and survival.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Yongxiao Yang, Zhengmei Li, Qin Li, Kai Ma, Yan Lin, Huigen Feng, Tianyun Wang
Summary: In this study, the expression of recombinant adalimumab in CHO cells was achieved through vector design and production process optimization, leading to increased yields of the monoclonal antibody.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Agronomy
Samuel T. Partey, Naresh V. Thevathasan, Robert B. Zougmore, Richard F. Preziosi
AGROFORESTRY SYSTEMS
(2018)
Article
Soil Science
Samuel T. Partey, Robert B. Zougmore, Naresh Thevathasan, Richard F. Preziosi
Article
Psychology, Biological
Christina R. Stanley, Huw Liddiard Williams, Richard F. Preziosi
Article
Ecology
Sarah M. Griffiths, Xavier A. Harrison, Che Weldon, Michael D. Wood, Abigail Pretorius, Kevin Hopkins, Graeme Fox, Richard F. Preziosi, Rachael E. Antwis
Review
Oceanography
Steven W. J. Canty, Richard F. Preziosi, Jennifer K. Rowntree
OCEAN & COASTAL MANAGEMENT
(2018)
Article
Biotechnology & Applied Microbiology
Maximilian Lularevic, Andrew J. Racher, Colin Jaques, Alexandros Kiparissides
BIOTECHNOLOGY AND BIOENGINEERING
(2019)
Article
Ecology
Sarah M. Griffiths, Rachael E. Antwis, Luca Lenzi, Anita Lucaci, Donald C. Behring, Mark J. Butler, Richard F. Preziosi
JOURNAL OF ANIMAL ECOLOGY
(2019)
Article
Biochemistry & Molecular Biology
Graeme Fox, Richard F. Preziosi, Rachael E. Antwis, Milena Benavides-Serrato, Fraser J. Combe, W. Edwin Harris, Ian R. Hartley, Andrew C. Kitchener, Selvino R. de Kort, Anne-Isola Nekaris, Jennifer K. Rowntree
MOLECULAR ECOLOGY RESOURCES
(2019)
Article
Agriculture, Dairy & Animal Science
Giovanni Quintavalle Pastorino, Richard Preziosi, Massimo Faustini, Giulio Curone, Mariangela Albertini, Dawn Nicoll, Lorna Moffat, Romain Pizzi, Silvia Mazzola
Article
Cell Biology
Jane F. Povey, Emily Saintas, Adewale V. Aderemi, Florian Rothweiler, Richard Zehner, Wilhelm G. Dirks, Jindrich Cinatl, Andrew J. Racher, Mark N. Wass, C. Mark Smales, Martin Michaelis
Article
Biotechnology & Applied Microbiology
James D. Budge, Tanya J. Knight, Jane Povey, Joanne Roobol, Ian R. Brown, Gurdeep Singh, Andrew Dean, Sarah Turner, Colin M. Jaques, Robert J. Young, Andrew J. Racher, C. Mark Smales
METABOLIC ENGINEERING
(2020)
Article
Pharmacology & Pharmacy
Joachim Schuster, Hanns-Christian Mahler, Atanas Koulov, Susanne Joerg, Andy Rachel, Joerg Huwyler, Pascal Detampel, Roman Mathaes
EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS
(2020)
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
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)
Article
Multidisciplinary Sciences
James D. Budge, Tanya J. Knight, Jane Povey, Joanne Roobol, Ian R. Brown, Gurdeep Singh, Andrew Dean, Sarah Turner, Colin M. Jaques, Robert J. Young, Andrew J. Racher, C. Mark Smales