Application of a fluorescent dye ‐based microfluidic sensor for real‐time detection of mAb aggregates
AbstractThe lack of process analytical technologies able to provide real-time information and process control over a biopharmaceutical process has long impaired the transition to continuous biomanufacturing. For the monoclonal antibody (mAb) production, aggregate formation is a major critical quality attribute (CQA) with several known process parameters (i.e., protein concentration and agitation) influencing this phenomenon. The development of a real-time tool to monitor aggregate formation is then crucial to gain control and achieve a continuous processing. Due to an inherent short operation time, miniaturized biosensors ...
Source: Biotechnology Progress - May 10, 2023 Category: Biotechnology Authors: Mariana N. S ão Pedro,
Michel H. M. Eppink,
Marcel Ottens Tags: RESEARCH ARTICLE Source Type: research
Predictive modeling of single pass tangential flow filtration for continuous biomanufacturing
AbstractOpportunities for process intensification have made continuous biomanufacturing an area of active research. While tangential flow filtration (TFF) is typically employed within the biologics purification train to increase drug substance concentration, single-pass TFF (SPTFF) modifies its format by enabling continuity of this process and achieving a multifold concentration factor through a single-pass over the filtration membranes. In continuous processes feed concentration and flow rate are determined by the preceding unit operations. Therefore, tight control of SPTFF output concentration must be achieved through pr...
Source: Biotechnology Progress - May 9, 2023 Category: Biotechnology Authors: Madeline Fuchs,
Rajan Bhawnani,
Sobhana A. Sripada,
Jessica Molek,
Mehdi Ghodbane Tags: RESEARCH ARTICLE Source Type: research
Understanding the effect of temperature downshift on CHO cell growth, antibody titer and product quality by intracellular metabolite profiling and in vivo monitoring of redox state
AbstractThe strategy of temperature downshift has been widely used in the biopharmaceutical industry to improve antibody production and cell-specific production rate (qp) with Chinese hamster ovary cells (CHO). However, the mechanism of temperature-induced metabolic rearrangement, especially important intracellular metabolic events, remains poorly understood. In this work, in order to explore the mechanisms of temperature-induced cell metabolism, we systematically assessed the differences in cell growth, antibody expression, and antibody quality between high-producing (HP) and low-producing (LP) CHO cell lines under both c...
Source: Biotechnology Progress - May 5, 2023 Category: Biotechnology Authors: Ziyu Zhu,
Xiaoqian Chen,
Wenhao Li,
Yingping Zhuang,
Yuzheng Zhao,
Guan Wang Tags: RESEARCH ARTICLE Source Type: research
Transcriptomic features reveal molecular signatures associated with recombinant adeno ‐associated virus production in HEK293 cells
This study investigated the transcriptomic features of two cell lines cultured in their respective media by comparing viral-producing cultures with non-producing cultures over time in parental human embryonic kidney cells (HEK293). The results demonstrate that the innate immune response signaling pathways of host cells (e.g., RIG-I-like receptor signaling pathway, Toll-like receptor signaling pathway, cytosolic DNA sensing pathway, JAK-STAT signaling pathway) were significantly enriched and upregulated. This was accompanied by the host cellular stress responses, including endoplasmic reticulum stress, autophagy, and apopto...
Source: Biotechnology Progress - May 3, 2023 Category: Biotechnology Authors: Yongdan Wang,
Qiang Fu,
Yong Suk Lee,
Sha Sha,
Seongkyu Yoon Tags: RESEARCH ARTICLE Source Type: research
CHO synthetic promoters improve expression and product quality of biotherapeutic proteins
AbstractWhen expressing complex biotherapeutic proteins, traditional expression plasmids and methods may not always yield sufficient levels of high-quality product. High-strength viral promoters commonly used for recombinant protein (rProtein) production in mammalian cells allow for maximal expression, but provide limited scope to alter their transcription dynamics. However, synthetic promoters designed to provide tunable transcriptional activity offer a plasmid engineering approach to more precisely regulate product quality, yield or to reduce product related contaminants. We substituted the viral promoter CMV with synthe...
Source: Biotechnology Progress - April 29, 2023 Category: Biotechnology Authors: Si Nga Sou,
Claire L. Harris,
Rebecca Williams,
Dorota Kozub,
Fabio Zurlo,
Yash D. Patel,
Praveen Kallamvalli Illam Sankaran,
Olalekan Daramola,
Adam Brown,
David C. James,
Diane Hatton,
Sarah Dunn,
Suzanne J. Gibson Tags: RESEARCH ARTICLE Source Type: research
Considerations for operational space definition and optimization of a no ‐salt flowthrough hydrophobic interaction chromatography purification step
AbstractNo-salt flowthrough hydrophobic interaction chromatography (HIC) has been shown to effectively remove process and product-related impurities from bioprocess streams. In this publication, a panel of six antibodies has been used to demonstrate operating principles for the application of no-salt flowthrough HIC in antibody purification processes. The results indicate that no-salt flowthrough HIC provides robust aggregate clearance across operating conditions including flow rate, and variations in resin ligand density. Additionally, HMW reduction has an optimal pH range relative to the isoelectric point of each molecul...
Source: Biotechnology Progress - April 28, 2023 Category: Biotechnology Authors: Allyson Tucker Dilks,
Joanne Gilchrist,
Yik Lam,
Nathan Nicholes,
Brad Stanley Tags: RESEARCH ARTICLE Source Type: research
An adaptive modeling approach using spiking ‐augmentation method to improve chemometric model performance in bioprocess monitoring
In this study, we developed an adaptive modeling strategy: the model was initially built using a calibration set of available perfusate and CB samples and then updated by augmenting spiking samples of the new molecules to the calibration set to make the model robust against perfusate or CB harvest of the new molecule. This strategy substantially improved the model performance and significantly reduced the modeling effort for new molecules. (Source: Biotechnology Progress)
Source: Biotechnology Progress - April 27, 2023 Category: Biotechnology Authors: Yuxiang Zhao,
Yawen Tang,
Dhanuka Wasalathanthri,
Jianlin Xu,
Julia Ding Tags: RESEARCH ARTICLE Source Type: research
