Article
Biotechnology & Applied Microbiology
Liang Zhang, MingLiang Wang, Andreas Castan, Hakan Hjalmarsson, Veronique Chotteau
Summary: This article discusses the importance of glycosylation in therapeutic monoclonal antibodies and validates the predictive ability of the modeling approach using Bayesian networks. Principal component analysis is shown to efficiently reduce input dimensions for glycoprotein production. The study demonstrates that a Bayesian network model can be a powerful tool in upstream process and medium development for glycoprotein productions.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Biotechnology & Applied Microbiology
Matthias Noebel, Craig Barry, Michael A. Macdonald, Kym Baker, Evan Shave, Stephen Mahler, Trent Munro, Veronica S. Martinez, Lars K. Nielsen, Esteban Marcellin
Summary: Chinese Hamster Ovary (CHO) cells have been increasingly used in biopharmaceutical production, particularly in perfusion culture mode. However, most current cell lines have been optimized for fed-batch culture methods and may not perform well under perfusion conditions. In this study, a triple knockout CHO cell line deficient in three apoptosis related genes was cultured in a perfusion system to improve cell resilience and viability. After an initial growth arrest phase, the cells entered a second growth phase, although with reduced productivity. Proteomics analysis revealed a correlation between growth arrest and cell cycle arrest, as well as upregulation of central carbon metabolism and oxidative phosphorylation. This upregulation was partially reversed during the second growth phase, suggesting intragenerational adaptations to stress. Our findings shed light on metabolic regulation and the potential development of better cell lines for perfusion cultivation.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
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
Young Hoon Oh, Kerri M. Mendola, Leila H. Choe, Lie Min, Ashton R. Lavoie, Sobhana A. Sripada, Taufika Islam Williams, Kelvin H. Lee, Yinges Yigzaw, Alexander Seay, Jerome Bill, Xuanwen Li, David J. Roush, Steven M. Cramer, Stefano Menegatti, Abraham M. Lenhoff
Summary: This study investigates the characterization and persistence of host-cell proteins (HCPs) in downstream processing steps for monoclonal antibody (mAb) manufacturing. The results show variability in HCP identities during processing steps, but consistency in the most abundant HCPs across different processes. The abundance of HCPs is likely related to the reproducibility of quantification measurements, and certain groups of HCPs may hinder characterization. The study suggests that factors such as HCP abundance and mAb-HCP interactions contribute to the persistence of individual HCPs and the identification of common, persistent HCPs in mAb manufacturing.
BIOTECHNOLOGY AND BIOENGINEERING
(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
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)
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)
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
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
Engineering, Chemical
Jan Mueller, Vivian Ott, Noemi Weiss, Peter Neubauer, Dieter Eibl, Regine Eibl
Summary: This study established a one-step inoculum production method based on an ultra-high cell density working cell bank, which can effectively improve the efficiency of producing biopharmaceuticals and increase manufacturing capacity.
CHEMIE INGENIEUR TECHNIK
(2022)
Article
Biotechnology & Applied Microbiology
Xiao-yin Wang, Qiu-jie Du, Wei-li Zhang, Dan-hua Xu, Xi Zhang, Yan-long Jia, Tian-yun Wang
Summary: This study investigated the effects of different poly A elements on transgene expression in CHO cells. The results showed that SV40 and Synt poly A significantly increased eGFP transgene expression and maintained long-term expression, while reducing expression variation. The choice of suitable poly A element is helpful in improving the expression of recombinant protein.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Nader Hashemi, Sayed Hassan Tabatabaee, Forough Shams, Azam Rahimpour, Bahram Kazemi, Masoumeh Rajabibazl, Javad Ranjbari
Summary: This study investigated the impact of SIRT6 overexpression on apoptosis-related gene expression, cell viability, apoptosis, and mAb productivity in CHO-K1 cells. The results showed that SIRT6 overexpression can improve cell viability, decrease apoptosis, and increase the expression of anti-CD52 IgG1 mAb.
MOLECULAR BIOLOGY REPORTS
(2023)
Article
Biotechnology & Applied Microbiology
Katrin Braasch, Marko Kryworuchko, James M. Piret
Summary: The modulation of autophagy has shown to increase productivity in CHO cell cultures, with the addition of AIP leading to a concentration-dependent enhancement in protein yield. However, this also results in a temporary drop in cell viability.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Review
Chemistry, Applied
Alana C. Szkodny, Kelvin H. Lee
Summary: This review highlights the key milestones in the production of biopharmaceuticals and emphasizes the maturity of the technologies used and the importance of understanding the interaction between product quality and processing conditions.
ANNUAL REVIEW OF CHEMICAL AND BIOMOLECULAR ENGINEERING
(2022)