Article
Biochemical Research Methods
Fang Zhao, Yuxiang Wan, Lei Nie, Jingyu Jiao, Dong Gao, Yan Sun, Zhenhua Chen, Yingting Shi, Jiayu Yang, Jianyang Pan, Haibin Wang, Haibin Qu
Summary: In this study, a workflow combining H-1 NMR and multivariate data analysis (MVDA) was developed for consistency monitoring and biochemical marker identification in commercial-scale CHO cell culture. The study identified a total of 63 metabolites in the CHO cell-free supernatants using H-1 NMR spectra. Multivariate statistical process control (MSPC) charts were used to evaluate process consistency, and orthogonal partial least square discriminant analysis (OPLS-DA) was used for biochemical marker identification during different cell growth phases. The combination of MVDA tools and H-1 NMR technology has great potential in biomanufacturing process research and can provide guidance for consistency evaluation and biochemical marker monitoring in the production of other biologics.
BIOTECHNOLOGY JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Zhe Lang, Shaofan Yan, Qingqing Xiong, Gong Chen
Summary: This study aimed to evaluate the feasibility of using a rocking type bioreactor system, specifically the WAVE 25, in an intensified perfusion culture mode for monoclonal antibody production in Chinese hamster ovary cell line. By using a disposable perfusion bag with a floating membrane and an automated filter switching system, the overall cell culture performance, product titer, and quality were compared to a typical IPC conducted in a glass bioreactor.
BIOTECHNOLOGY LETTERS
(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)
Article
Biotechnology & Applied Microbiology
Sophie Maria, Laurent Bonneau, Benjamin Fould, Gilles Ferry, Jean Albert Boutin, Charlotte Cabanne, Xavier Santarelli, Gilles Joucla
Summary: In this study, three culture methods were compared to determine the perfusion process parameters for a CHO cell line producing a monoclonal antibody. The push-to-low (PTL) and perfusion-without-bleeding (PWB) methods resulted in lower CSPRmin estimations, suggesting a three-step approach to determine the perfusion parameters.
BIOCHEMICAL ENGINEERING JOURNAL
(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
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
Jianlin Xu, Shun Zheng, Zeinab Dawood, Charles Hill, Weixin Jin, Xuankuo Xu, Julia Ding, Michael C. Borys, Sanchayita Ghose, Zheng Jian Li, Girish Pendse
Summary: The research aimed to develop intensified processes for the production of a monoclonal antibody, resulting in improved productivity but facing product quality challenges. Introducing carboxypeptidase B treatment effectively improved product quality and increased yield, demonstrating potential application for other processes with similar issues.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
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
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
Magdalena Pappenreiter, Benjamin Bayer, Marijan Logarusic, Bernhard Sissolak, Alois Jungbauer
Summary: The study investigated the individual impacts of mild and rigorous intra-experimental parameter shifts on the culture performance and cell metabolism of an antibody producing CHO cell line. The results showed significant changes in cell growth, metabolism, and productivity due to intra-experimental parameter shifts. However, if the conditions within an experiment are changed, rigorous parameter shifts should be avoided, as they can lead to irreversible changes in cell behavior, as observed in media blending experiments.
BIOCHEMICAL ENGINEERING JOURNAL
(2023)
Article
Engineering, Chemical
Heribert Helgers, Axel Schmidt, Jochen Strube
Summary: The development of new biologics is becoming more challenging due to global competition and increased requirements for process understanding and quality assurance. Predictive and mechanistic process models are needed to reduce resources and time in process development and enable automated process control.
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
Biotechnology & Applied Microbiology
Dennis Weber, Christian Sittig, Juergen Hubbuch
Summary: Freezing of cell culture supernatant in mAb platform process development helps concentrate certain proteins and particles, leading to better removal of host cell proteins during purification steps. The freezing temperature also affects the concentrations of HCP and mAb dimers in the eluate during protein A chromatography. Rapid freezing of CCS in process development is crucial for maintaining comparability and transferability to manufacturing.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
Article
Multidisciplinary Sciences
Qin He, Jianlin Xu, Erik Vandermark, Jun Tian, Yueming Qian, Cynthia Chelius, Jongchan Lee, Michael C. Borys, Zheng Jian Li, Girish Pendse
Summary: This study utilized high-throughput screening (HTS) to identify a chemically defined media for late phase process development, resulting in a two-fold increase in titer. It demonstrates the potential of HTS as an effective approach for determining media composition and supplementation.
Article
Biochemical Research Methods
Iman Shahidi Pour Savizi, Ehsan Motamedian, Nathan E. Lewis, Ioscani Jimenez del Val, Seyed Abbas Shojaosadati
Summary: Monoclonal antibodies (mABs) are important therapeutic proteins whose immunogenicity and efficacy depend on the glycosylation process, which can be influenced by bioprocess conditions. Ammonium has been shown to induce heterogeneity in protein glycosylation by altering the sialic acid content. Three mechanisms have been proposed to explain how this process is influenced by ammonium.
BIOTECHNOLOGY JOURNAL
(2021)
Review
Biotechnology & Applied Microbiology
Jan Niklas Pauk, Janani Raju Palanisamy, Julian Kager, Krisztina Koczka, Gerald Berghammer, Christoph Herwig, Lukas Veiter
Summary: Overexpression of recombinant proteins in Escherichia coli can lead to the formation of inclusion bodies, but these are now being viewed as a promising production method. Refolding is necessary to obtain the correct active protein product, but current technology faces challenges like buffer usage, mixing uniformity, and low re-folding yields. Quality by Design methods, online monitoring techniques, and digital twin technology may offer a solution to these challenges.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Peter Sinner, Marlene Stiegler, Oliver Goldbeck, Gerd M. Seibold, Christoph Herwig, Julian Kager
Summary: Model-based state estimators enable online monitoring of bioprocesses, taking into account the variability of metabolic capacities. This study presents a particle filtering framework that continuously adapts physiological parameters by Bayesian inference.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Chemistry, Analytical
Katharina Boettinger, Wolfgang Esser-Skala, Marius Segl, Christoph Herwig, Christian G. Huber
Summary: The study used a simple HPLC-MS workflow to analyze mAb product quality in real-time at different fermentation time points. The results revealed the presence of misassembled variants in the fermentation broth, alongside changes in mAb glycosylation profile over time, with galactosylated variants being the most abundant. The workflow proved to be robust for analyzing the complex matrix with low standard deviations.
ANALYTICA CHIMICA ACTA
(2022)
Article
Biochemical Research Methods
Patrick Romann, Jakub Kolar, Daniela Tobler, Christoph Herwig, Jean-Marc Bielser, Thomas K. Villiger
Summary: This study developed a systematic Raman calibration workflow for perfusion processes in biopharmaceutical processes. The method allows for fast and specific model calibration, and can be applied to monitor other analytes of interest.
BIOTECHNOLOGY JOURNAL
(2022)
Article
Biotechnology & Applied Microbiology
Thomas Oberleitner, Thomas Zahel, Barbara Pretzner, Christoph Herwig
Summary: This study aims to develop a new experimental design method that gains information about final product quality by minimizing the simulated out-of-specification rate. The proposed method uses a chain of regression models to map process parameters to critical quality attributes for each unit operation and has been tested in different scenarios.
BIOENGINEERING-BASEL
(2022)
Article
Biotechnology & Applied Microbiology
Patrick Romann, Jakub Kolar, Loic Chappuis, Christoph Herwig, Thomas K. Villiger, Jean-Marc Bielser
Summary: Bleed recycling is a novel method for increasing the yield of steady-state perfusion processes. By concentrating process bleed and selectively removing biomass, it allows for significant product saving. The use of inclined gravity settling as a bleed recycling technology showed similar efficiency to acoustic separation and had no negative impact on cell viabilities and product quality. With reduced system complexity and scalability, inclined gravity settling is the preferred technology for perfusion processes.
BIOCHEMICAL ENGINEERING JOURNAL
(2023)
Article
Computer Science, Interdisciplinary Applications
Don Fabian Muller, Daniel Wibbing, Christoph Herwig, Julian Kager
Summary: In this study, we propose a soft sensor that accurately estimates the yield coefficient Y-x/s and the substrate uptake capacity q(Smax) in a cultivation process. The estimation is achieved through offgas measurements and a nonlinear state observer with an underlying mechanistic model. Various simulation scenarios were analyzed to determine estimation accuracy under uncertain measurements. In-silico testing showed that q(Smax) can be estimated with reasonable accuracy depending on the sensitivity of parameter Y-x/s. The developed state and parameter estimation were verified in induced Escherichia coli cultivations, allowing for accurate prediction of biomass and substrate accumulation, as well as detection of decreasing Y-x/s and q(Smax).
COMPUTERS & CHEMICAL ENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
M. Adnan Jouned, Julian Kager, Vignesh Rajamanickam, Christoph Herwig, Tilman Barz
Summary: Intermittent bolus feeding in minibioreactor systems has a significant impact on the metabolism of E. coli cultivations. The temporary surplus of substrates and transient oxygen limitation caused by bolus feeding triggers the production of inhibitory byproducts. This study explores the dissolved oxygen tension (DOT) response in E. coli minibioreactor cultivations and identifies a unique response behavior in the DOT signal that is related to a certain dilution ratio per biomass unit. The analysis suggests a plausible relationship between metabolic adaptation and the newly observed DOT signal segment, which has not been reported in previous literature. Hypotheses linking specific DOT segments to specific metabolic states are proposed, and quantitative analysis and mechanistic model simulations support these hypotheses, showing the possibility of extracting cell physiological and growth parameters from the DOT signal.
BIOENGINEERING-BASEL
(2023)
Article
Biotechnology & Applied Microbiology
Oscar Fabian Garcia Aponte, Bence Kozma, Dominik Egger, Cornelia Kasper, Christoph Herwig
Summary: Understanding the growth and functionality variation of lymphocytes during expansion is crucial for process control and endpoint determination. However, offline cell characterization hinders timely identification of process deviations and transitions. In this study, a culturing protocol was developed to investigate the day-to-day variation of NK-92 cells' cytotoxicity, revealing two distinctive phases with exponential growth and intermittent drop in viability. Biomass volume was found to be correlated with cytotoxicity, providing potential performance indicators for non-invasive monitoring of CQAs.
BIOCHEMICAL ENGINEERING JOURNAL
(2023)
Article
Biochemical Research Methods
Patrick Romann, Sebastian Schneider, Daniela Tobler, Martin Jordan, Arnaud Perilleux, Jonathan Souquet, Christoph Herwig, Jean-Marc Bielser, Thomas K. Villiger
Summary: The Raman-controlled pyruvate feeding strategy stabilized metabolic activity and prevented product quality drifting in perfusion cell cultures.
BIOTECHNOLOGY JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Lukas Marschall, Chitti Babu Gottimukkala, Biswajit Kayal, Veerabhadra Madurai Veeraraghavan, Samir Kumar Mandal, Suman Bandyopadhyay, Christoph Herwig
Summary: Temperature downshift is a crucial control strategy in mammalian cell culture processes, which can prolong production phases and enhance productivity. However, for the development of biosimilars, achieving a specified product quality is the main challenge. In this study, we investigated the impact of temperature shifts on the glycosylation profile of a monoclonal antibody (mAb) and found that an upshift in temperature to 37 degrees C can optimize the glycosylation profile. The optimized control strategy was successfully implemented at the manufacturing scale, demonstrating the potential of temperature upshifts in improving the quality of other mAbs.
BIOENGINEERING-BASEL
(2023)
Article
Biotechnology & Applied Microbiology
Patrick Romann, Philip Giller, Antony Sibilia, Christoph Herwig, Andrew L. Zydney, Arnaud Perilleux, Jonathan Souquet, Jean-Marc Bielser, Thomas K. Villiger
Summary: This study investigated the benefits of co-current filtrate flow in hollow fiber-based membrane filtration. The results showed that co-current flow can improve product sieving and alleviate common challenges in hollow fiber-based systems. Therefore, co-current flow represents an interesting and scalable alternative in tangential flow filtration (TFF) applications.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Christopher Taylor, Barbara Pretzner, Thomas Zahel, Christoph Herwig
Summary: Integrated process models can serve as the engine of a digital asset in a multistep-process digital twin, but face limitations in their deployment. This study improves integrated process models by introducing a simplified data model, increasing statistical robustness, adding a new simulation flow for scale-dependent variables, and describing a novel algorithm for data-driven environments. The proposed architectural and procedural requirements for a deployed digital twin provide a final framework for a digital asset in bioprocessing.
BIOENGINEERING-BASEL
(2022)
Article
Biotechnology & Applied Microbiology
Rudiger W. Maschke, Barbara Pretzner, Gernot T. John, Christoph Herwig, Dieter Eibl
Summary: Shake flasks are widely used in biotechnology, but they lack online data recording and control. This study evaluates the feasibility of using online data to determine key performance indicators (KPIs) by comparing different online measurement methods. The results show that online data can effectively determine KPIs in shake flask cultivations, providing guidance for subsequent process development.
BIOENGINEERING-BASEL
(2022)
Article
Biotechnology & Applied Microbiology
Thomas Mainka, Christoph Herwig, Stefan Pflugl
Summary: This study investigates the influence of critical process parameters on the performance of a continuous treatment process for residual process brine using halophilic microorganisms. The study proposes optimized operating conditions for efficient degradation of organics in the residual process brine.
FERMENTATION-BASEL
(2022)