Demonstration of a robust high cell density transient CHO platform yielding mAb titers of up to 2 g/L without medium exchange
In this study, we have demonstrated a simple high-titer platform for TGE in CHO based on a dilution process of seeding cells. We achieved titers ranging from 0.8 to 1.9 g/L for eight model mAbs at three scales (1, 30, 100 mL) in 10 days using our new platform. The ability to seed by dilution significantly streamlined the process and dramatically enhanced platform throughput. We observed a modest reduction in titer ranging from 11% to 28% when cells were seed ed using dilution compared to when cells were seeded using medium exchange. Further studies revealed that carry over of spent medium into transfection negativel...
Source: Biotechnology Progress - February 9, 2024 Category: Biotechnology Authors: Rigumula Wu,
Danielle M. Kahl,
Ronald Kloberdanz,
Kushal J. Rohilla,
Sowmya Balasubramanian Tags: RESEARCH ARTICLE Source Type: research
Implementation of mDoE ‐methods to a microcarrier‐based expansion processes for mesenchymal stem cells
In this study, the mDoE workflow was used for the development of an expansion processes with human immortalized mesenchymal stem cells (hMSC-TERT) and the aim of maximizing cell yield assuming only a limited amount of prior knowledge at a very early stage of development. First, suitable microcarriers for expansion in shake flasks were screened and the differentiation of the cells was proven. Second, initial experiments were performed to generate prior knowledge, which was then used to set up the mathematical model and to estimate the model parameters. Finally, the mDoE was used to determine and evaluate the design space to...
Source: Biotechnology Progress - February 9, 2024 Category: Biotechnology Authors: Kim B. Kuchem üller,
Ralf Pörtner,
Johannes Möller Tags: RESEARCH ARTICLE Source Type: research
Demonstration of a robust high cell density transient CHO platform yielding mAb titers of up to 2 g/L without medium exchange
In this study, we have demonstrated a simple high-titer platform for TGE in CHO based on a dilution process of seeding cells. We achieved titers ranging from 0.8 to 1.9 g/L for eight model mAbs at three scales (1, 30, 100 mL) in 10 days using our new platform. The ability to seed by dilution significantly streamlined the process and dramatically enhanced platform throughput. We observed a modest reduction in titer ranging from 11% to 28% when cells were seed ed using dilution compared to when cells were seeded using medium exchange. Further studies revealed that carry over of spent medium into transfection negativel...
Source: Biotechnology Progress - February 8, 2024 Category: Biotechnology Authors: Rigumula Wu,
Danielle M. Kahl,
Ronald Kloberdanz,
Kushal J. Rohilla,
Sowmya Balasubramanian Tags: RESEARCH ARTICLE Source Type: research
Effect of flux and shear rate on E. coli recovery in tangential flow filtration through a single hollow fiber
AbstractPathogenic bacteria which enter a viable but non-culturable (VBNC) state impede efforts to reach detectable concentrations required for PCR methods. This motivated a strategy for tangential flow filtration to concentrate bacteria in aqueous samples while maintaining the bacteria in a viable state, maximizing their recovery and achieving high fluxes through a single hollow fiber membrane. Filtrations were carried out for green fluorescent protein (GFP)E. coli at high shear rates (up to 27,000 sec−1) through 0.2 μm cut-off polyethersulfone (PES) microfilter membranes or 50 kDa polysulfone (PS) ultrafilter ...
Source: Biotechnology Progress - February 8, 2024 Category: Biotechnology Authors: Jessica Zuponcic,
Fernanda Cunha,
Grant Springer,
Eduardo Ximenes,
Michael R. Ladisch Tags: RESEARCH ARTICLE Source Type: research
Gene copy number, gene configuration and LC/HC mRNA ratio impact on antibody productivity and product quality in targeted integration CHO cell lines
AbstractThe augmentation of transgene copy numbers is a prevalent approach presumed to enhance transcriptional activity and product yield. CHO cell lines engineered via targeted integration (TI) offer an advantageous platform for investigating the interplay between gene copy number, mRNA abundance, product yield, and product quality. Our investigation revealed that incrementally elevating the gene copy numbers of both IgG heavy chain (HC) and light chain (LC) concurrently resulted in the attainment of plateaus in mRNA levels and product titers, notably occurring beyond four to five gene copies integrated at the same TI sit...
Source: Biotechnology Progress - February 7, 2024 Category: Biotechnology Authors: Zion Lee,
Jun Wan,
Amy Shen,
Gavin Barnard Tags: RESEARCH ARTICLE Source Type: research
Enhancing cell separation in a hybrid spiral dielectrophoretic microchannel: Numerical insights and optimal operating conditions
AbstractReliable separation of circulating tumor cells from blood cells is crucial for early cancer diagnosis and prognosis. Many conventional microfluidic platforms take advantage of the size difference between particles for their separation, which renders them impractical for sorting overlapping-sized cells. To address this concern, a hybrid inertial-dielectrophoretic microfluidic chip is proposed in this work for continuous and single-stage separation of lung cancer cell line A549 cells from white blood cells of overlapping size. The working mechanism of the proposed spiral microchannel embedded with planar interdigitat...
Source: Biotechnology Progress - January 31, 2024 Category: Biotechnology Authors: Mohammed Raihan Uddin,
Xiaolin Chen Tags: RESEARCH ARTICLE Source Type: research
Evaluating end ‐to‐end continuous antibody manufacture with column‐free capture alternatives from economic, environmental, and robustness perspectives
This article describes a decisional tool that encompasses mass balance and design equations, process economics, stochastic simulation and multi-criteria decision-making and enables the evaluation of different batch, and continuous flowsheets for monoclonal antibody (mAb) manufacture. The traditional batch process was compared with end-to-end continuous bioprocesses with either protein A capture or column-free capture employing aqueous two-phase extraction or precipitation from economic, environmental, and robustness perspectives. The cost of goods analysis predicted that continuous flowsheets could offer substantial cost s...
Source: Biotechnology Progress - January 30, 2024 Category: Biotechnology Authors: Catarina P. G. Neves,
Jonathan L. Coffman,
Suzanne S. Farid Tags: RESEARCH ARTICLE Source Type: research
Multi ‐omics kinetic analysis of recombinant adeno‐associated virus production by plasmid transfection of HEK293 cells
AbstractRecombinant adeno-associated virus (rAAV) is among the most commonly used vectors for gene therapy. It is commonly produced by transfection of HEK293 cells with three plasmids each containing the vector genome including gene of interest (GOI), helper functions, andrep andcap genes for genome replication and capsid formation. To meet the potential clinical needs, the productivity of the production system needs to be enhanced. A better process characterization of the production system will further advance our insights into ways to enhance productivity. Here, we employed transcriptomic analysis to quantify the dynamic...
Source: Biotechnology Progress - January 30, 2024 Category: Biotechnology Authors: Min Lu,
Zion Lee,
Wei ‐Shou Hu Tags: RESEARCH ARTICLE Source Type: research
Process development and characterization for integrated continuous bioprocesses —Highlights from N‐mAb
AbstractThe N-mAb case study was produced by the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) to support teaching and learning for both industry and to accelerate adoption of advanced manufacturing process technologies such as integrated continuous bioprocesses (ICB) for mAbs. Similar to the A-mAb case study, N-mAb presents the evolution of an integrated control strategy, from early clinical through process validation and commercial manufacturing with a focus on elements that are unique to integrated continuous bioprocesses. This publication presents a summary of the process design and cha...
Source: Biotechnology Progress - January 30, 2024 Category: Biotechnology Authors: Kevin Brower,
Kelly Wiltberger,
Claudia Berdugo,
Allen Bosley,
Elizabeth Goodrich,
Valerie Pferdeort,
Gene Schaefer Tags: RESEARCH ARTICLE Source Type: research
