Modeling scalability of impurity precipitation in downstream biomanufacturing
The objective of this work is to compare isoelectric impurity precipitation behavior for a monoclonal antibody product across scales, from benchtop to pilot manufacturing. Scaling parameters such as agitation and vessel geometry were investigated, with the precipitate amount and particle size distribution (PSD) characterized via turbidity and flow imaging microscopy. Qualitative analysis of the data shows that maintaining a consistent energy dissipation rate (EDR) could be used for approximate scaling of vessel geometry and agitator speeds in the absence of more detailed simulation. For a more rigorous approach, however, a...
Source: Biotechnology Progress - March 28, 2024 Category: Biotechnology Authors: Jing Guo,
Steven J. Traylor,
Mohamed Agoub,
Weixin Jin,
Helen Hua,
R. Bertrum Diemer,
Xuankuo Xu,
Sanchayita Ghose,
Zheng Jian Li,
Abraham M. Lenhoff Tags: RESEARCH ARTICLE Source Type: research
Raman spectroscopy and one ‐dimensional convolutional neural network modeling as a real‐time monitoring tool for in vitro transaminase‐catalyzed synthesis of a pharmaceutically relevant amine precursor
AbstractRaman spectroscopy has been used to measure the concentration of a pharmaceutically relevant model amine intermediate for positive allosteric modulators of nicotinic acetylcholine receptor in a ω-transaminase-catalyzed conversion. A model based on a one-dimensional convolutional neural network was developed to translate raw data augmented Raman spectra directly into substrate concentrations, with which the conversion from ketone to amine by ω-transaminase could be determined over time. T he model showed very good predictive capabilities, withR2 values higher than 0.99 for the spectra included in the modeling and ...
Source: Biotechnology Progress - March 27, 2024 Category: Biotechnology Authors: Julie Østerby Madsen,
Sebastian Olivier Nymann Topalian,
Mikkel Fog Jacobsen,
Tommy Skovby,
Krist V. Gernaey,
Allan S. Myerson,
John Woodley Tags: RESEARCH ARTICLE Source Type: research
Development of a high ‐throughput scale‐down model in Ambr® 250 HT for plasmid DNA fermentation processes
AbstractRecent advances in messenger ribonucleic acid (mRNA) vaccines and gene therapy vectors have increased the need for rapid plasmid DNA (pDNA) screening and production within the biopharmaceutical industry. High-throughput (HT) fermentor systems, such as the Ambr ® 250 HT, can significantly accelerate process development timelines of pDNA upstream processes compared to traditional bench-scale glass fermentors or small-scale steam-in-place (SIP) fermentors. However, such scale-down models must be qualified to ensure that they are representative of the larger scale process similar to traditional small-scale models. In ...
Source: Biotechnology Progress - March 18, 2024 Category: Biotechnology Authors: Shu Fang,
Dillon J. Sinanan,
Marc H. Perez,
Ra úl G. Cruz‐Quintero,
Sachin R. Jadhav Tags: RESEARCH ARTICLE Source Type: research
A modular and multi ‐functional purification strategy that enables a common framework for manufacturing scale integrated and continuous biomanufacturing
This study utilized an Apollo™ X CHO-DG44 mAb-expressing cell line in a steady-state perfusion bioreactor, harvesting product continuously with a cell retention device and connected SymphonX™ purification skids. The dow nstream process used the same chemistry (resins, buffer composition, membrane composition) as our historical batch processing platform, with SymphonX™ in-line conditioning and buffer concentrates. We used surge vessels between unit operations, single-column chromatography (protein A, cation and an ion exchange) and two-tank batch virus inactivation. After the first polishing step (cation exchange), we...
Source: Biotechnology Progress - March 18, 2024 Category: Biotechnology Authors: Leon P. Pybus,
Charles Heise,
Tibor Nagy,
Carmen Heeran,
Terri Dover,
John Raven,
Junichi Kori,
Graeme Burton,
Hiroshi Sakuyama,
Benjamin Hastings,
Michelle Lyons,
Shinichi Nakai,
Jonathan Haigh Tags: RESEARCH ARTICLE Source Type: research
Methods comparison of two ‐dimensional gel electrophoresis for host cell protein characterization
AbstractTwo-dimensional electrophoresis (2DE) is a gel-based protein separation method based on size and charge which is commonly used for the characterization of host cell proteins (HCPs) during drug development in biotech and pharmaceutical companies. HCPs are a heterogenous mixture of proteins produced by host cells during a biologics drug manufacturing process. Different gel electrophoresis methods including traditional 2D SDS-PAGE with silver and SYPRO Ruby fluorescent dye staining as well as two-dimensional difference gel electrophoresis (2D-DIGE) were compared for their relative abilities to characterize HCPs. SYPRO...
Source: Biotechnology Progress - March 18, 2024 Category: Biotechnology Authors: Abigail King,
Yiwei Zhao,
Alexandru Lazar,
Margeaux Capron,
Niranjan Thiruvur,
Xinrong Liu Tags: RESEARCH ARTICLE Source Type: research
Bioburden detection on surface and water samples in a rapid, ultra ‐sensitive and high‐throughput manner
The objective of this study is to develop and validate an assay and protocol for detecting bioburden on solid surfaces, as well as in water, with high sensitivity and accuracy in a rapid manner. Henceforth, a resazurin-based assay optimized for detecting bioburden has been integrated with a previously developed portable multichannel fluorometer. The microbes were isolated from solid surfaces in different laboratory settings by swabbing technique, and stream water was collected for contamination analysis. Based on the results, the assay and protocol can successfully detect bioburden as low as 20 CFU/cm2 and 10 CFU/mL ...
Source: Biotechnology Progress - March 18, 2024 Category: Biotechnology Authors: Md Sadique Hasan,
Chad Sundberg,
Elias Gilotte,
Xudong Ge,
Yordan Kostov,
Govind Rao Tags: RESEARCH ARTICLE Source Type: research
Proteomic analysis of host cell protein fouling during bioreactor harvesting
AbstractChinese hamster ovary (CHO) cells are among the most common cell lines used for therapeutic protein production. Membrane fouling during bioreactor harvesting is a major limitation for the downstream purification of therapeutic proteins. Host cell proteins (HCP) are the most challenging impurities during downstream purification processes. The present work focuses on identification of HCP foulants during CHO bioreactor harvesting using reverse asymmetrical commercial membrane BioOptimal ™ MF-SL. In order to investigate foulants and fouling behavior during cell clarification, for the first time a novel backwash proc...
Source: Biotechnology Progress - March 14, 2024 Category: Biotechnology Authors: Da Zhang,
S. Ranil Wickramasinghe,
Andrew L. Zydney,
John P. Smelko,
Abdullah Loman,
April Wheeler,
Xianghong Qian Tags: RESEARCH ARTICLE Source Type: research
When will we have a clone? An industry perspective on the typical CLD timeline
AbstractCell line development (CLD) represents a complex but highly critical process during the development of a biological drug. To shed light on this crucial workflow, a team of BioPhorum members (authors) has developed and executed surveys focused on the activities and effort involved in a typical CLD campaign. An average of 27 members from different companies that participate in the BioPhorum CLD working group answered surveys covering three distinguishable stages of a standard CLD process: (1) Pre-transfection, including vector design and construction; (2) Transfection, spanning the initial introduction of vector into...
Source: Biotechnology Progress - March 13, 2024 Category: Biotechnology Authors: Howard Clarke,
Anke Mayer ‐Bartschmid,
Chenxing Zheng,
Elizabeth Masterjohn,
Falguni Patel,
Mark Moffat,
Qingxiang Wei,
Ren Liu,
Robyn Emmins,
Simon Fischer,
Stephanie Rieder,
Thomas Kelly Tags: REVIEW Source Type: research
Host cell proteins in monoclonal antibody processing: Control, detection, and removal
AbstractHost cell proteins (HCPs) are process-related impurities in a therapeutic protein expressed using cell culture technology. This review presents biopharmaceutical industry trends in terms of both HCPs in the bioprocessing of monoclonal antibodies (mAbs) and the capabilities for HCP clearance by downstream unit operations. A comprehensive assessment of currently implemented and emerging technologies in the manufacturing processes with extensive references was performed. Meta-analyses of published downstream data were conducted to identify trends. Improved analytical methods and understanding of “high-risk” HCPs l...
Source: Biotechnology Progress - March 13, 2024 Category: Biotechnology Authors: Takao Ito,
Herb Lutz,
Lihan Tan,
Bin Wang,
Janice Tan,
Masum Patel,
Lance Chen,
Yuki Tsunakawa,
Byunghyun Park,
Subhasis Banerjee Tags: REVIEW Source Type: research
Measurement and control of foam generation in a mammalian cell culture
This study showed the value of an optical monitoring system (LabCam) for measuring foam generation and dissipation in a bioreactor to assess th e efficiency of antifoam agents to reduce foam in a bioreactor. This has the potential for use as a control system that could be designed for continuous monitoring and foam control in a mammalian cell bioprocess. (Source: Biotechnology Progress)
Source: Biotechnology Progress - March 13, 2024 Category: Biotechnology Authors: James Flynn,
Laura Breen,
Shankara Narayanan,
Michael Butler Tags: RESEARCH ARTICLE Source Type: research
Surface decoration of low molecular weight polyethylenimine (LMW PEI) by phthalated dextrin for improved delivery of interleukin ‐12 plasmid
AbstractIn this investigation, low molecular weight polyethyleneimine (LMW PEI; 1.8 kDa branched PEI) was conjugated to phathalated dextrin. The aim of this chemical modification was to decorate PEI molecules with a hydrophilic layer to improve its biophysical properties while the phthalic moiety may improve the hydrophilic-hydrophobic balance of the final structure. The polymer s were prepared at various conjugation degrees ranging from 6.5% to 16.5% and characterized in terms of biophysical characteristics as well as their gene transfer ability and cell-induced toxicity. The results showed that dextrin-phthalated-PEI ...
Source: Biotechnology Progress - March 11, 2024 Category: Biotechnology Authors: Valiollah Keshavarz,
Maryam Kazemi,
Bahman Khalvati,
Fateme Zare,
Ali Dehshahri,
Hossein Sadeghpour Tags: RESEARCH ARTICLE Source Type: research
Simplifying stable CHO cell line generation with high probability of monoclonality by using microfluidic dispensing as an alternative to fluorescence activated cell sorting
AbstractSingle cell cloning is a critical step for cell line development (CLD) for therapeutic protein production, with proof of monoclonality being compulsorily sought in regulatory filings. Among the different single cell deposition technologies, we found that fluorescence activated cell sorting (FACS) offers high probability of monoclonality and can allow selective enrichment of the producer cells. However, FACS instruments are expensive and resource-intensive, have a large footprint, require highly skilled operators and take hours for setup, thereby complicating the cell line generation process. With the aim of finding...
Source: Biotechnology Progress - March 11, 2024 Category: Biotechnology Authors: Lina Chakrabarti,
James Savery,
John Patrick Mpindi,
Judith Klover,
Lina Li,
Jie Zhu Tags: RESEARCH ARTICLE Source Type: research
High ‐yield recombinant adeno‐associated viral vector production by multivariate optimization of bioprocess and transfection conditions
AbstractRecombinant adeno-associated viral vectors (rAAVs) are one of the most used vehicles for gene therapy, with five rAAV therapeutics commercially approved by the FDA. To improve product yield, we optimized the suspension production process of rAAV8 vectors carrying a proprietary transgene using a commercially available transfection reagent, FectoVIR-AAV. Using a miniaturized automated 250 mL scale bioreactor system, we generated models of vector genome (vg) titer, capsid (cp) titer, and Vg:Cp percentage from two multivariate design of experiment studies, one centered around bioreactor operating parameters, and ano...
Source: Biotechnology Progress - March 7, 2024 Category: Biotechnology Authors: Louis Coplan,
Zhe Zhang,
Nicole Ragone,
John Reeves,
Audrey Rodriguez,
Aishwarya Shevade,
Hanne Bak,
Andrew D. Tustian Tags: RESEARCH ARTICLE Source Type: research
Integration of rapid bioburden testing into production quality management systems and process control
AbstractThe move to integrated continuous bioprocessing (ICB), while providing a means for process intensification, can put added strain on process analytics when conventional methods are used. For instance, traditional microbial methods provide minimal value to ICB processes given that the time required for data to become available is much longer than a typical full cycle of the manufacturing process. Although rapid microbial detection has been in discussion for over 30 years, it is still not routinely deployed in commercial biopharmaceutical manufacturing. One contributing factor is the ability to integrate this techn...
Source: Biotechnology Progress - March 7, 2024 Category: Biotechnology Authors: Irina Ramos,
Michelle Najera,
Gene Schaefer Tags: REVIEW Source Type: research
Leveraging bioanalytical characterization of fractionated monoclonal antibody pools to identify aggregation ‐prone and less filterable proteoforms during virus filtration
AbstractMonoclonal antibodies (mAbs) are an essential class of biotherapeutics. A platform process is used for mAb development to ensure clinically safe and stable molecules. Regulatory authorities ensure that mAb production processes include sufficient viral clearance steps to achieve less than one virus particle per million doses of product. Virus filtration is used for size-based removal of enveloped and nonenveloped viruses during downstream processing of mAbs. Process development in mAb purification relies on empirical approaches and often includes adsorptive prefiltration to mitigate virus filter fouling. Opportuniti...
Source: Biotechnology Progress - March 7, 2024 Category: Biotechnology Authors: Solomon Isu,
Lilia Vinskus,
Derek Silva,
Kristina Cunningham,
Thomas Elich,
Patricia Greenhalgh,
Adam Sokolnicki,
Bala Raghunath Tags: RESEARCH ARTICLE Source Type: research
