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Methods comparison of two-dimensional gel electrophoresis for host cell protein characterization Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-18 Abigail King, Yiwei Zhao, Alexandru Lazar, Margeaux Capron, Niranjan Thiruvur, Xinrong Liu
Two-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
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A modular and multi-functional purification strategy that enables a common framework for manufacturing scale integrated and continuous biomanufacturing Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-18 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
Biopharmaceutical manufacture is transitioning from batch to integrated and continuous biomanufacturing (ICB). The common framework for most ICB, potentially enables a global biomanufacturing ecosystem utilizing modular and multi-function manufacturing equipment. Integrating unit operation hardware and software from multiple suppliers, complex supply chains enabled by multiple customized single-use
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Development of a high-throughput scale-down model in Ambr® 250 HT for plasmid DNA fermentation processes Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-18 Shu Fang, Dillon J. Sinanan, Marc H. Perez, Raúl G. Cruz-Quintero, Sachin R. Jadhav
Recent 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
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Bioburden detection on surface and water samples in a rapid, ultra-sensitive and high-throughput manner Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-17 Md Sadique Hasan, Chad Sundberg, Elias Gilotte, Xudong Ge, Yordan Kostov, Govind Rao
Bioburden detection is crucial for food, water, and biopharmaceutical applications as it can directly impact public health. 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
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Host cell proteins in monoclonal antibody processing: Control, detection, and removal Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-13 Takao Ito, Herb Lutz, Lihan Tan, Bin Wang, Janice Tan, Masum Patel, Lance Chen, Yuki Tsunakawa, Byunghyun Park, Subhasis Banerjee
Host 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
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When will we have a clone? An industry perspective on the typical CLD timeline Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-13 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
Cell 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
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Proteomic analysis of host cell protein fouling during bioreactor harvesting Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-13 Da Zhang, S. Ranil Wickramasinghe, Andrew L. Zydney, John P. Smelko, Abdullah Loman, April Wheeler, Xianghong Qian
Chinese 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
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Measurement and control of foam generation in a mammalian cell culture Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-13 James Flynn, Laura Breen, Shankara Narayanan, Michael Butler
Foam is generated in mammalian cell cultures by excessive agitation or gas sparging. This occurs particularly in cultures that generate recombinant proteins at high cell concentrations. Three antifoam agents were tested for their compatibility with antibody‐producing Chinese hamster ovary (CHO) cells. One agent (antifoam 204) was completely inhibitory to growth at a concentration of 10 ppm, one agent
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Surface decoration of low molecular weight polyethylenimine (LMW PEI) by phthalated dextrin for improved delivery of interleukin‐12 plasmid Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-11 Valiollah Keshavarz, Maryam Kazemi, Bahman Khalvati, Fateme Zare, Ali Dehshahri, Hossein Sadeghpour
In 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 polymers were prepared at various conjugation
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Simplifying stable CHO cell line generation with high probability of monoclonality by using microfluidic dispensing as an alternative to fluorescence activated cell sorting Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-11 Lina Chakrabarti, James Savery, John Patrick Mpindi, Judith Klover, Lina Li, Jie Zhu
Single 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
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Leveraging bioanalytical characterization of fractionated monoclonal antibody pools to identify aggregation‐prone and less filterable proteoforms during virus filtration Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-07 Solomon Isu, Lilia Vinskus, Derek Silva, Kristina Cunningham, Thomas Elich, Patricia Greenhalgh, Adam Sokolnicki, Bala Raghunath
Monoclonal 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
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Integration of rapid bioburden testing into production quality management systems and process control Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-07 Irina Ramos, Michelle Najera, Gene Schaefer
The 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
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High‐yield recombinant adeno‐associated viral vector production by multivariate optimization of bioprocess and transfection conditions Biotechnol. Prog. (IF 2.9) Pub Date : 2024-03-07 Louis Coplan, Zhe Zhang, Nicole Ragone, John Reeves, Audrey Rodriguez, Aishwarya Shevade, Hanne Bak, Andrew D. Tustian
Recombinant 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
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Catechins prevent monoclonal antibody fragmentation during production via fed‐batch culture of Chinese hamster ovary cells Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-28 Tsuyoshi Yamaguchi, Hiroko Ishikawa, Mie Fukuda, Yumi Sugita, Misaki Furuie, Ryuma Nagano, Toshiyuki Suzawa, Koichi Yamamoto, Kaori Wakamatsu
Chinese hamster ovary (CHO) cells are widely used for the industrial production of therapeutic monoclonal antibodies (mAbs). To meet the increasing market demands, high productivity, and quality are required in cell culture. One of the critical attributes of mAbs, from a safety perspective, is mAb fragmentation. However, methods for preventing mAbs fragmentation in CHO cell culture are limited. In
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Application of fucosylation inhibitors for production of afucosylated antibody Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-28 Ping Xu, Yu Chuan Ou, Michael Smith, Jim Paulson, Michael A. Schmidt, Lakshmi Kandari, Rodney Parsons, Anurag Khetan
Fucosylation is an important quality attribute for therapeutic antibodies. Afucosylated antibodies exhibit higher therapeutic efficacies than their fucosylated counterparts through antibody‐dependent cellular cytotoxicity (ADCC) mechanism. Since higher potency is beneficial in reducing dose or duration of the treatment, afucosylated antibodies have attracted a great deal of interest in biotherapeutics
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Fed‐batch performance profiles for mAb production using different intensified N − 1 seed strategies are CHO cell‐line dependent Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-28 Yawen Tang, Jianlin Xu, Mengmeng Xu, Zhuangrong Huang, Johanna Santos, Qin He, Michael Borys, Anurag Khetan
Recent optimizations of cell culture processes have focused on the final seed scale‐up step (N − 1 stage) used to inoculate the production bioreactor (N‐stage bioreactor) to enable higher inoculation cell densities (2–20 × 106 cells/mL), which could shorten the production culture duration and/or increase the volumetric productivity. N − 1 seed process intensification can be achieved by either non‐perfusion
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Biotransformation of maize bran‐derived ferulic acid to vanillin using an adapted strain of Amycolatopsis sp. ATCC 39116 Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-28 Rasika V. Tupe, Nitesh K. Singh, Annamma A. Odaneth
Maize bran, an agro‐processing waste residue, is a good source of ferulic acid that can be further valorized for vanillin production. However, extraction of ferulic acid from natural sources has been challenging due to low concentrations and intensive extraction procedures. In the present work, ferulic acid streams (purities ranging from 5% to 75%) extracted from maize bran using thermochemical methods
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A deep‐well plate enabled automated high‐throughput cell line development platform Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-20 Xiaoyan Tang, Jorge Quiroz, Yixiao Zhang, Jessica Pan, Zhong Lai, Zhimei Du, Ren Liu
Cell line development (CLD) plays a crucial role in the manufacturing process development of therapeutic biologics. Most biologics are produced in Chinese hamster ovary (CHO) cell. Because of the nature of random transgene integration in CHO genome and CHO's inherent plasticity, stable CHO transfectants usually have a vast diversity in productivity, growth, and product quality. Thus, we often must
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Evaluation of PDL1 positive cancer cell‐specific binding activity of recombinant anti‐PDL1 scFv Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-20 Sun‐Hee Kim, Hae‐Min Park, Hee‐Jin Jeong
Programmed cell death‐ligand 1 (PDL1) is a transmembrane protein that is characterized as an immune regulatory molecule. We recently developed a recombinant single‐chain fragment of variable domain (scFv) against PDL1, which showed high binding efficiency to purified recombinant PDL1 protein. However, at that time, proof‐of‐concept data for the effect of scFv using PDL1‐expressing cells was lacking
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Effect of inner diameter, filter length, and pore size on hollow fiber filter fouling during perfusion cell culture Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-11 Dominique WuDunn, Andrea Squeri, Jimmy Vu, Ashna Dhingra, Jon Coffman, Ken Lee
As the need for higher volumetric productivity in biomanufacturing grows, biopharmaceutical companies are increasingly investing in a perfusion cell culture process, most commonly one that uses a hollow fiber filter as the cell retention device. A current challenge with using hollow fiber filters is fouling of the membrane, which reduces product sieving and can increase transmembrane pressure (TMP)
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Advancing multiproduct resin reuse for development and clinical manufacturing of an antibody-based therapeutic Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-09 Hong Li, Patricia Rose, Patricia Rowicki, Collette Cutler, Jeffrey T. McPhee, Claudia Frey, Linda Lemieux, Gerald Pelette, Joo Kok Ang, Ren Liu, Douglas D. Richardson
Chromatography resins used for purifying biopharmaceuticals are generally dedicated to a single product. For clinical manufacturing, this can result in resin being used only for a fraction of its potential lifetime. Extending the use of resins to multiple products can significantly reduce resin waste and cost. It can also improve manufacturing flexibility in case of raw material shortage during times
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Implementation of mDoE-methods to a microcarrier-based expansion processes for mesenchymal stem cells Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-09 Kim B. Kuchemüller, Ralf Pörtner, Johannes Möller
The need for advanced therapy medicinal products (ATMPs) has gained increased attention in recent years. In this respect, a well-designed cell expansion process is needed to efficiently manufacture the required number of cells with the desired product quality. This step is challenging due to the biological complexity of the respective primary cell (e.g., mesenchymal stem cells (MSC)) and the usage
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Demonstration of a robust high cell density transient CHO platform yielding mAb titers of up to 2 g/L without medium exchange Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-08 Rigumula Wu, Danielle M. Kahl, Ronald Kloberdanz, Kushal J. Rohilla, Sowmya Balasubramanian
Biopharmaceuticals like therapeutic monoclonal antibodies (mAbs) and other derived proteins are popular for treating various diseases. Transient gene expression (TGE) is typically used as a fast yet efficient method to generate moderate amounts of material. It has been used to support early stage research and discovery processes. Introduction of a robust high yielding and predictive TGE platform in
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Gene copy number, gene configuration and LC/HC mRNA ratio impact on antibody productivity and product quality in targeted integration CHO cell lines Biotechnol. Prog. (IF 2.9) Pub Date : 2024-02-06 Zion Lee, Jun Wan, Amy Shen, Gavin Barnard
The 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
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Process development and characterization for integrated continuous bioprocesses—Highlights from N-mAb Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-30 Kevin Brower, Kelly Wiltberger, Claudia Berdugo, Allen Bosley, Elizabeth Goodrich, Valerie Pferdeort, Gene Schaefer
The 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,
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Evaluating end-to-end continuous antibody manufacture with column-free capture alternatives from economic, environmental, and robustness perspectives Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-30 Catarina P. G. Neves, Jonathan L. Coffman, Suzanne S. Farid
Process intensification efforts have renewed interest in the potential of end-to-end continuous manufacture with column-free capture alternatives. 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
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Development of bioreactor scale-down model using orthogonal projections to latent structures method and CO2 supplementation Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-30 Jinxin Gao, Laurie B. Hazeltine, Neal Stroud, Ning Liu, Yao-ming Huang
Scale-down model qualification is an important step for developing a large-scale cell culture process to enhance process understanding and support process characterization studies. Traditionally, only harvest data are used to show consistency between small-scale and large-scale bioreactor performance, allowing attributes that are dynamic over the cell culture period to be overlooked. A novel statistical
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Multi-omics kinetic analysis of recombinant adeno-associated virus production by plasmid transfection of HEK293 cells Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-30 Min Lu, Zion Lee, Wei-Shou Hu
Recombinant 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, and rep and cap genes for genome replication and capsid formation. To meet the potential clinical needs, the productivity of the production
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Enhancing cell separation in a hybrid spiral dielectrophoretic microchannel: Numerical insights and optimal operating conditions Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-30 Mohammed Raihan Uddin, Xiaolin Chen
Reliable 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
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Correction to “Investigation of the hepatic respiration and liver zonation on rat hepatocytes using an integrated oxygen biosensor in a microscale device” Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-30
Satomi Matsumoto, Astia R. Safitri, Matheu Danoy, Toshiro Maekawa, Haruyuki Kinoshita, Marie Shinohara, Yasuyuki Sakai, Teruo Fujii, Eric Leclerc. In the third row of Figure 8, images for APC, PCK1, and DAPI at the inlet were the same as the ones at the outlet. Also, the images for Betacatenin were taken from another biological replicate than APC, PCK1 or DAPI. FIGURE 8 Open in figure viewerPowerPoint
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Transitioning from static to suspension culture system for large-scale production of xeno-free extracellular vesicles derived from mesenchymal stromal cells Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-21 Natália Cristine Dias dos Santos, Paula Bruzadelle-Vieira, Nádia de Cássia Noronha, Amanda Mizukami-Martins, Maristela Delgado Orellana, Maria Vitória L. B. Bentley, Dimas Tadeu Covas, Kamilla Swiech, Kelen Cristina Ribeiro Malmegrim
Extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) have shown increasing therapeutic potential in the last years. However, large production of EV is required for therapeutic purposes. Thereby, scaling up MSC cultivation in bioreactors is essential to allow culture parameters monitoring. In this study, we reported the establishment of a scalable bioprocess to produce MSC-EV in
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A custom 3D printed paddlewheel improves growth in flat panel photobioreactor Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-21 Michelle Meagher, Jacob Tamburro, Nanette R. Boyle
One of the main challenges with using flat panel photobioreactors for algal growth is uneven mixing and settling of cells in corners, especially when bubbling is the only method used for mixing. In order to improve mixing in our flat panel reactor, we designed a custom paddlewheel. Paddlewheels are frequently used in outdoor algae raceway ponds to improve mixing and we are taking advantage of the same
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Control strategy for biopharmaceutical production by model predictive control Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-10 Touraj Eslami, Alois Jungbauer
The biopharmaceutical industry is rapidly advancing, driven by the need for cutting-edge technologies to meet the growing demand for life-saving treatments. In this context, Model Predictive Control (MPC) has emerged as a promising solution to address the complexity of modern biopharmaceutical production processes. Its ability to optimize operations and ensure consistent product yields has made it
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Adaptive modeling optimized by the data fusion strategy: Real-time dying cell percentage prediction using capacitance spectroscopy Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-04 Suyang Wu, Stephanie A. Ketcham, Claudia Corredor, Douglas Both, Yuxiang Zhao, James K. Drennen, Carl A. Anderson
The previous research showcased a partial least squares (PLS) regression model accurately predicting cell death percentages using in-line capacitance spectra. The current study advances the model accuracy through adaptive modeling employing a data fusion approach. This strategy enhances prediction performance by incorporating variables from the Cole–Cole model, conductivity and its derivatives over
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Chitosan based hybrid superabsorbent for controlled drug delivery application Biotechnol. Prog. (IF 2.9) Pub Date : 2024-01-03 Medha, Sapna Sethi
In the present study, a hybrid chitosan-alginate superabsorbent is prepared using maleic acid as a cross-linker and acrylamide as a grafting agent using the free radical mechanism. The composite hydrogel shows good swelling capacity along with hemocompatibility and biocompatibility and hence it is utilized as a drug delivery device. The characterization techniques including x-ray diffraction, Fourier
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Impedance nanobiosensor based on enzyme-conjugated biosynthesized gold nanoparticles for the detection of Gram-positive bacteria Biotechnol. Prog. (IF 2.9) Pub Date : 2023-12-31 Sarani Sen, Priyabrata Sarkar
In this report, gold nanoparticles (GNPS) were synthesized using cell-free extracts of seven different isolates, namely, Pseudomonas aerogenosa CEBP2, Pseudomonas sp. CEBP1, Pseudomonas pseudoalcaligenes CEB1G, Acinetobactor baumani CEBS1, Cuprividus sp. CEB3, Micrococcus luteus CUB12, and Pandoraea sp. CUB2S. The spectroscopic (UV–vis, FTIR, DLS, XRD, EDS) and microscopic (FESEM, TEM) results confirm
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Adapting virus filtration to continuous processing: Effects of product and process variability on filtration performance Biotechnol. Prog. (IF 2.9) Pub Date : 2023-12-25 Julie Kozaili, William Rayfield, Adrian Gospodarek, Mark Brower, Daniel Strauss
Virus filtration (VF) is an important unit operation in the manufacture of biotherapeutics that provides robust removal of potential virus contaminants. Small virus removal can be impacted by the low operating pressures and potential depressurization events that are often associated with continuous operations where increased operational flexibility for higher loading at low flux and low pressure is
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Effect of pH, NaCl concentration, and mAb concentration of feed solution on the filterability of Planova™ 20N and Planova™ BioEX Biotechnol. Prog. (IF 2.9) Pub Date : 2023-12-25 Chie Hashikawa-Muto, Yoshihiro Yokoyama, Ryo Hamamoto, Kazuya Kobayashi, Yumiko Masuda, Koichi Nonaka
Virus filtration is one of the most important steps in ensuring viral safety during the purification of monoclonal antibodies (mAbs) and other biotherapeutics derived from mammalian cell cultures. Regarding the various virus retentive filters, including Planova filters, a great deal of data has been reported on the virus retention capability and its mechanism. Along with the virus retention capability
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Difficult-to-express antigen generation through a co-expression and disassociation methodology Biotechnol. Prog. (IF 2.9) Pub Date : 2023-12-13 Ricky Lieu, Grace Chao, Emma Kennedy, J. Michael Sauder, Prabakaran Narayanasamy, Anna Pustilnik, Adithi Thangaraju, Carolyn Ho, Mariah J. Pedroza, Diana Ruiz, Xiaomin Yang
Extracellular domain (ECD) antigens are crucial components for antibody discovery, in vitro assays, and epitope mapping during therapeutical antibody development. Oftentimes, those antigens are difficult to produce while retaining the biologic function/activity upon extracellular secretion in commonly used expression systems. We have developed an effective method to cope with the challenge of generating
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Exploring preferred binding domains of IgG1 mAbs to multimodal adsorbents using a combined biophysics and simulation approach Biotechnol. Prog. (IF 2.9) Pub Date : 2023-12-03 Kabir Dhingra, Imee Sinha, Mark Snyder, David Roush, Steven M. Cramer
In this work, we employ a recently developed biophysical technique that uses diethylpyrocarbonate (DEPC) covalent labeling and mass spectrometry for the identification of mAb binding patches to two multimodal cation exchange resins at different pH. This approach compares the labeling results obtained in the bound and unbound states to identify residues that are sterically shielded and thus located
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Managing integrated continuous bioprocesses in real time: Deviations in product quality Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-28 Gustavo Grampp, Allen Bosley, Maen Qadan, John Schiel, Andy Spasoff, Pascal Valax, Gene Schaefer
The 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 regulators around adoption of advanced manufacturing process technologies such as integrated continuous bioprocesses (ICB) for monoclonal antibodies (mAbs). N-mAb presents the evolution of an integrated control strategy, from
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An automated high inoculation density fed-batch bioreactor, enabled through N-1 perfusion, accommodates clonal diversity and doubles titers Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-28 Mikayla Olin, Nicolas Wolnick, Hunter Crittenden, Anthony Quach, Brian Russell, Shannon Hendrick, Julia Armstrong, Thaddaeus Webster, Brian Hadley, Marissa Dickson, Jessica Hodgkins, Kevin Busa, Roger Connolly, Brandon Downey
An important consideration for biopharmaceutical processes is the cost of goods (CoGs) of biotherapeutics manufacturing. CoGs can be reduced by dramatically increasing the productivity of the bioreactor process. In this study, we demonstrate that an intensified process which couples a perfused N-1 seed reactor and a fully automated high inoculation density (HID) N stage reactor substantially increases
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Intensified perfusion culture (IPC) reduced recombinant protein fragmentation Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-24 Ge Yan, Xun Lu, Ruiqiang Sun, Weichang Zhou, Hang Zhou
Mammalian cells remain the mainstay of biological production host. In industry, cultivating and harvest strategies are sorted in batch mode (e.g., batch, fed-batch, concentrated fed-batch and intensified fed-batch) and continuous mode (e.g., perfusion). To retrieve greater productivity and better product quality, especially for the sensitive products prone to fragmentation, culture modes with various
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Understanding adsorption behavior of Polysorbate-20 to sterile filters in therapeutic proteins final filtration process Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-21 Jian Ren, Yun Zhang, Chen Wang
Surfactants are commonly used in the therapeutic protein manufacturing process as stabilizer. Polysorbate-20 (PS-20) is one of the most commonly used surfactants to mitigate protein aggregation in the therapeutic protein formulation. It has been observed that polysorbate can be adsorbed by sterile filters during the final filtration process, which poses risk of uneven distribution and potentially reduced
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Model validation and selection in metabolic flux analysis and flux balance analysis Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-24 Joshua A. M. Kaste, Yair Shachar-Hill
13C-Metabolic Flux Analysis (13C-MFA) and Flux Balance Analysis (FBA) are widely used to investigate the operation of biochemical networks in both biological and biotechnological research. Both methods use metabolic reaction network models of metabolism operating at steady state so that reaction rates (fluxes) and the levels of metabolic intermediates are constrained to be invariant. They provide estimated
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Dynamic flux balance analysis of 1,3-propanediol production by clostridium butyricum fermentation Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-20 Duo-Tao Pan, Pan Wang, Xiao-Li Wang, Ya-Qin Sun, Zhi-Long Xiu
To study the relationship between the yield of 1,3-propanediol (1,3-PDO) and the flux change of the Clostridium butyricum metabolic pathway, an optimized calculation method based on dynamic flux balance analysis was used by combining genome-scale flux balance analysis with a kinetic model. A more comprehensive and extensive metabolic pathway was obtained by optimization calculations. The primary extended
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Integration of Planova filters in manufacturing processes of biologicals improve the virus safety effectively: A review of publicly available data Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-20 Albrecht Gröner
The capacity to remove viruses by Planova filters produced by Asahi Kasei, primarily by small virus-retentive filters, were compiled from data in peer-reviewed publications and, partly, publicly available data from presentations at conferences (Planova workshops). Data from more than 100 publications and presentations at conferences covering Planova filters were assessed. The data were grouped according
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Effect of adenosine and cordycepin on recombinant antibody production yields in two different Chinease hamster ovary cell lines Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-20 Salinthip Jarusintanakorn, Kuntalee Rangnoi, Montarop Yamabhai, Enrico Mastrobattista
In this study, we investigated the effect of adenosine and its derivative cordycepin on the production yield of a recombinant human monoclonal antibody (adalimumab) in two commonly used Chinese Hamster ovary (CHO) cell lines that have different gene amplification systems, namely CHO-DHFR− and GS-CHO knockout (GS-KO CHO) cells and that were grown in batch culture, with and without glucose feeding. The
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Depth filtration for clarification of intensified lentiviral vector suspension cell culture Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-20 Mukesh Mayani, Srinivas Nellimarla, Ratish Mangalathillam, Hema Rao, Susannah Patarroyo-White, Junfen Ma, Bruno Figueroa
Depth filtration significantly impacts efficiency of lentiviral (LV) vector purification process. However, it is often deprioritized in the overall scope of viral vector manufacturing process optimization. The demand for LV vectors has increased with the rise in disease indications, making it crucial to improve current manufacturing processes. Upstream bioreactor process intensification has enabled
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Polyhydroxyalkanoates and exopolysaccharides: An alternative for valuation of the co-production of microbial biopolymers Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-20 Edmilson Clarindo de Siqueira, Aline de Andrade Alves, Páblo Eugênio da Costa e Silva, Maria Paloma Silva de Barros, Laureen Michelle Houllou
Polyhydroxyalkanoates (PHAs) and exopolysaccharides (EPSs) belong to a class of abundant biopolymers produced by various fermenting microorganisms. These biocompounds have high value-added potential and can be produced concurrently. Co-production of PHAs and EPSs is a strategy employed by researchers to reduce costs associated with large-scale production. EPSs and PHAs are non-toxic, biocompatible
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Impact of interstitial flow on cartilage matrix synthesis and NF-kB transcription factor mRNA expression in a novel perfusion bioreactor Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-20 Haneen A. Abusharkh, Terreill Robertson, Juana Mendenhall, Bulent A. Gozen, Edwin M. Tingstad, Nehal I. Abu-Lail, David B. Thiessen, Bernard J. Van Wie
This work is focused on designing an easy-to-use novel perfusion system for articular cartilage (AC) tissue engineering and using it to elucidate the mechanism by which interstitial shear upregulates matrix synthesis by articular chondrocytes (AChs). Porous chitosan-agarose (CHAG) scaffolds were synthesized and compared to bulk agarose (AG) scaffolds. Both scaffolds were seeded with osteoarthritic
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Efficiency of ultrafiltration/diafiltration in removing organic and elemental process equipment related leachables from biological therapeutics Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-14 Bin Sun, Mahsa Hadidi, Jose Santiago Nuñez, Benben Song, Gilbert E. Tumambac, Ken Wong, Gregor Kalinowski, James J. Hathcock
In the production of biological therapeutics such as monoclonal antibodies (mAbs), ultrafiltration and diafiltration (UF/DF) are widely regarded as effective downstream processing steps capable of removing process equipment related leachables (PERLs) introduced upstream of the UF/DF step. However, clearance data available in the literature are limited to species with low partition coefficients (log P)
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Removal of phenolic inhibitors from lignocellulose hydrolysates using laccases for the production of fuels and chemicals Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-14 M. T. Fernández-Sandoval, A. García, K. V. Teymennet-Ramírez, D. Y. Arenas-Olivares, F. Martínez-Morales, M. R. Trejo-Hernández
Lignocellulose is the most abundant biopolymer in the biosphere. It is inexpensive and therefore considered an attractive feedstock to produce biofuels and other biochemicals. Thermochemical and/or enzymatic pretreatment is used to release fermentable monomeric sugars. However, a variety of inhibitory by-products such as weak acids, furans, and phenolics that inhibit cell growth and fermentation are
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Correlation of microbial community structure and volatile flavor compounds during corn yellow wine fermentation Biotechnol. Prog. (IF 2.9) Pub Date : 2023-11-13 Jianhua Guo, Xiaoxu Zhao, Jie Shi
High-throughput sequencing was used to define microbial community structure and GC–MS to identify volatile flavor substances during fermentation of corn yellow wine, and results were analyzed by multivariate statistical analysis. Seventeen bacterial phyla, 239 bacterial genera, 4 fungal phyla, and 18 fungal genera were found and changes in community structure occurred during fermentation. Twenty-four
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Proliferation of mammalian cells with Chlorococcum littorale algal compounds without serum support Biotechnol. Prog. (IF 2.9) Pub Date : 2023-10-30 Jayeesha Ghosh, Yoshikatsu Akiyama, Yuji Haraguchi, Kumiko Yamanaka, Toru Asahi, Yoichi Nakao, Tatsuya Shimizu
In recent years, serum-free medium for mammalian cell cultivation has attracted a lot of attention, considering the high cost of production and environmental load involved in developing the conventional animal sera. The use of alternative growth-promoting products in mammalian cell cultivation such as extracts from microalgae has proven to be quite beneficial and environmental-friendly. This research
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Utilizing targeted integration CHO pools to potentially accelerate the GMP manufacturing of monoclonal and bispecific antibodies Biotechnol. Prog. (IF 2.9) Pub Date : 2023-10-24 Gavin C. Barnard, Michelle Zhou, Amy Shen, Inn H. Yuk, Michael W. Laird
Monoclonal antibodies (mAbs) are effective therapeutic agents against many acute infectious diseases including COVID-19, Ebola, RSV, Clostridium difficile, and Anthrax. mAbs can therefore help combat a future pandemic. Unfortunately, mAb development typically takes years, limiting its potential to save lives during a pandemic. Therefore “pandemic mAb” timelines need to be shortened. One acceleration
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Shear stress and microRNAs for better metastatic cancer management Biotechnol. Prog. (IF 2.9) Pub Date : 2023-10-16 Siluveru KrishnaPriya, Pallavi S. Nair, Prerna Bhalla, D. Karunagaran, G. K. Suraishkumar
Metastasis is the process by which cancer cells move from the primary location to establish themselves in a new location in the human body. It is still a significant challenge in cancer management because it is responsible for 90% of cancer-related deaths. In this work, we present an idea to use shear stress encountered by all metastasizing cells as an elegant means to deactivate metastasizing cancer
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Economic simulation of batch and continuous aqueous two-phase purification for viral products Biotechnol. Prog. (IF 2.9) Pub Date : 2023-10-16 Natalie M. Nold, Eric Pearson, Caryn L. Heldt
Vaccine manufacturing strategies that lower capital and production costs could improve vaccine access by reducing the cost per dose and encouraging localized manufacturing. Continuous processing is increasingly utilized to drive lower costs in biological manufacturing by requiring fewer capital and operating resources. Aqueous two-phase systems (ATPS) are a liquid–liquid extraction technique that enables
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A novel method for continuous chromatographic separation of monoclonal antibody charge variants by combining displacement mode chromatography and step elution Biotechnol. Prog. (IF 2.9) Pub Date : 2023-10-12 Anupa Anupa, Vikrant Bansode, Nikhil Kateja, Anurag S. Rathore
Charge heterogeneity of monoclonal antibodies is considered a critical quality attribute and hence needs to be monitored and controlled by the manufacturer. Typically, this is accomplished via separation of charge variants on cation exchange chromatography (CEX) using a pH or conductivity based linear gradient elution. Although an effective approach, this is challenging particularly during continuous