Biotechnology Progress最新文献_第9页

Proteomic analysis of host cell protein fouling during bioreactor harvesting 生物反应器收获过程中宿主细胞蛋白质污垢的蛋白质组分析。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-13 DOI: 10.1002/btpr.3453

Da Zhang, S. Ranil Wickramasinghe, Andrew L. Zydney, John P. Smelko, Abdullah Loman, April Wheeler, Xianghong Qian

{"title":"Proteomic analysis of host cell protein fouling during bioreactor harvesting","authors":"Da Zhang, S. Ranil Wickramasinghe, Andrew L. Zydney, John P. Smelko, Abdullah Loman, April Wheeler, Xianghong Qian","doi":"10.1002/btpr.3453","DOIUrl":"10.1002/btpr.3453","url":null,"abstract":"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 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 process was developed to effectively elute almost all the HCP and DNA from the fouled membrane filter. The isoelectric points (pIs) and molecular weights (MWs) of major HCP in the bioreactor harvest and fouled on the membrane were successfully characterized using two-dimensional gel electrophoresis (2D SDS-PAGE). In addition, a total of 8 HCP were identified using matrix-assisted laser desorption/ionization-mass spectroscopy (MALDI-MS). The majority of these HCP are enzymes or associated with exosomes, both of which can form submicron-sized particles which could lead to the plugging of the filters.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

When will we have a clone? An industry perspective on the typical CLD timeline 我们何时才能拥有克隆人？从行业角度看典型的 CLD 时间表。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-13 DOI: 10.1002/btpr.3449

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

{"title":"When will we have a clone? An industry perspective on the typical CLD timeline","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","doi":"10.1002/btpr.3449","DOIUrl":"10.1002/btpr.3449","url":null,"abstract":"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 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 cells and subsequent selection and analysis of the pools; and (3) Single Cell Cloning and Lead Clone Selection, comprising methods of isolating single cells and confirming clonal origin, subsequent expansion and screening processes, and methods for identifying and banking lead clones. The surveys were very extensive, including a total of 341 questions split between antibody and complex molecule CLD processes. In this survey review, the authors interpret and highlight responses for antibody development and, where relevant, contrast complex molecule development challenges to provide a comprehensive industry perspective on the typical time and effort required to develop a CHO production cell line.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3449","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140109000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Host cell proteins in monoclonal antibody processing: Control, detection, and removal 单克隆抗体加工过程中的宿主细胞蛋白：控制、检测和清除。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-13 DOI: 10.1002/btpr.3448

Takao Ito, Herb Lutz, Lihan Tan, Bin Wang, Janice Tan, Masum Patel, Lance Chen, Yuki Tsunakawa, Byunghyun Park, Subhasis Banerjee

{"title":"Host cell proteins in monoclonal antibody processing: Control, detection, and removal","authors":"Takao Ito, Herb Lutz, Lihan Tan, Bin Wang, Janice Tan, Masum Patel, Lance Chen, Yuki Tsunakawa, Byunghyun Park, Subhasis Banerjee","doi":"10.1002/btpr.3448","DOIUrl":"10.1002/btpr.3448","url":null,"abstract":"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 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 lead to more robust manufacturing processes and higher-quality therapeutics. The trend of higher cell density cultures leads to both higher mAb expression and higher HCP levels. However, HCP levels can be significantly reduced with improvements in operations, resulting in similar concentrations of approx. 10 ppm HCPs. There are no differences in the performance of HCP clearance between recent enhanced downstream operations and traditional batch processing. This review includes best practices for developing improved processes.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3448","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Measurement and control of foam generation in a mammalian cell culture 测量和控制哺乳动物细胞培养过程中泡沫的产生。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-12 DOI: 10.1002/btpr.3450

James Flynn, Laura Breen, Shankara Narayanan, Michael Butler

{"title":"Measurement and control of foam generation in a mammalian cell culture","authors":"James Flynn, Laura Breen, Shankara Narayanan, Michael Butler","doi":"10.1002/btpr.3450","DOIUrl":"10.1002/btpr.3450","url":null,"abstract":"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 (antifoam C) showed partial inhibition and a third (antifoam SE-15) showed no inhibition at this concentration. A novel foam image analyzer (LabCam) was used to evaluate two antifoams (C and SE-15) for their ability to dissipate foam generated in cell culture media by enhanced agitation. The presence of antifoam in the media reduced significantly the foam layer that was generated and this was shown to be rapidly dissipated in the presence of 10 ppm SE-15. The antifoams were also tested for foam dissipation in cultures of CHO cells at >106 cells/mL. Supplementation of the cultures with SE-15 resulted in dissipation of foam generated by excessive gas sparging within 2 min. Under equivalent conditions 75% of foam dissipated in the presence of antifoam C, within 2 min but there was a residual foam layer up to 25 min. This study showed the value of an optical monitoring system (LabCam) for measuring foam generation and dissipation in a bioreactor to assess the 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.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3450","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108998","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Simplifying stable CHO cell line generation with high probability of monoclonality by using microfluidic dispensing as an alternative to fluorescence activated cell sorting 利用微流体喷点技术替代荧光激活细胞分选技术，简化具有高单克隆概率的稳定 CHO 细胞系的生成。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-10 DOI: 10.1002/btpr.3441

Lina Chakrabarti, James Savery, John Patrick Mpindi, Judith Klover, Lina Li, Jie Zhu

{"title":"Simplifying stable CHO cell line generation with high probability of monoclonality by using microfluidic dispensing as an alternative to fluorescence activated cell sorting","authors":"Lina Chakrabarti, James Savery, John Patrick Mpindi, Judith Klover, Lina Li, Jie Zhu","doi":"10.1002/btpr.3441","DOIUrl":"10.1002/btpr.3441","url":null,"abstract":"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, 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 an easy-to-use alternative to FACS, we identified a flow cytometry-based microfluidic cell dispenser, which presents a single cell sorting solution for biopharmaceutical CLD. The microfluidic cell dispenser is small, budget-friendly, easy-to-use, requires lower-cost consumables, permits flow cytometry-enabled multiparametric target cell enrichment and offers fast and gentle single cell dispensing into multiwell plates. Following comprehensive evaluation, we found that single cell deposition by the microfluidic cell dispenser resulted in >99% probability of monoclonality for production cell lines. Moreover, the clonally derived producer cell lines generated from the microfluidic cell dispenser demonstrated comparable or improved growth profiles and production capability compared to the FACS derived cell lines. Taken together, microfluidic cell dispensing can serve as a cost-effective, efficient and convenient alternative to FACS, simplifying the biopharmaceutical CLD platform with significant reductions in both scientist time and running costs.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3441","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140093341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Surface decoration of low molecular weight polyethylenimine (LMW PEI) by phthalated dextrin for improved delivery of interleukin-12 plasmid 用邻苯二甲酸糊精对低分子量聚乙烯亚胺（LMW PEI）进行表面装饰，以改善白细胞介素-12 质粒的输送。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-10 DOI: 10.1002/btpr.3443

Valiollah Keshavarz, Maryam Kazemi, Bahman Khalvati, Fateme Zare, Ali Dehshahri, Hossein Sadeghpour

{"title":"Surface decoration of low molecular weight polyethylenimine (LMW PEI) by phthalated dextrin for improved delivery of interleukin-12 plasmid","authors":"Valiollah Keshavarz, Maryam Kazemi, Bahman Khalvati, Fateme Zare, Ali Dehshahri, Hossein Sadeghpour","doi":"10.1002/btpr.3443","DOIUrl":"10.1002/btpr.3443","url":null,"abstract":"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 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 (DPHPEI) polymer was able to form nanoparticles with the size range of around 118–170 nm, with the zeta potential of 6.2–9.5 mV. DPHPEI polymers could increase the level of desired protein expression in the cells by up to three folds compared with unmodified LMW PEI while the cell viability of the modified polymers was around 80%. The result of this study shows a promising approach to improve the transfection efficiency of LMW PEI while maintaining its low toxic effects.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140093342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Integration of rapid bioburden testing into production quality management systems and process control 将快速生物负载测试纳入生产质量管理系统和流程控制。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-07 DOI: 10.1002/btpr.3431

Irina Ramos, Michelle Najera, Gene Schaefer

{"title":"Integration of rapid bioburden testing into production quality management systems and process control","authors":"Irina Ramos, Michelle Najera, Gene Schaefer","doi":"10.1002/btpr.3431","DOIUrl":"10.1002/btpr.3431","url":null,"abstract":"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 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 technology into a process control strategy and existing quality systems. An understanding of the capability of microbial detection technology available today can be leveraged to implement a control strategy for bioburden monitoring in real time for process intermediates. One key tenet of this proposed control strategy is the use of a “two-tiered approach” wherein a fast (but possibly less sensitive) test is used to monitor the process and trigger further action for a second, longer duration test which is used to confirm and quantify the presence of bioburden and identify the organism. This approach, presented here alongside several case studies for microbial monitoring, can have broader application for other process analytical technologies where fit for purpose methods could be employed to establish process control alongside real time continuous processes.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3431","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140048667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-yield recombinant adeno-associated viral vector production by multivariate optimization of bioprocess and transfection conditions 通过对生物工艺和转染条件进行多元优化，实现高产重组腺相关病毒载体的生产。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-07 DOI: 10.1002/btpr.3445

Louis Coplan, Zhe Zhang, Nicole Ragone, John Reeves, Audrey Rodriguez, Aishwarya Shevade, Hanne Bak, Andrew D. Tustian

{"title":"High-yield recombinant adeno-associated viral vector production by multivariate optimization of bioprocess and transfection conditions","authors":"Louis Coplan, Zhe Zhang, Nicole Ragone, John Reeves, Audrey Rodriguez, Aishwarya Shevade, Hanne Bak, Andrew D. Tustian","doi":"10.1002/btpr.3445","DOIUrl":"10.1002/btpr.3445","url":null,"abstract":"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 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 another based on the transfection conditions. Using the optimized process returned from these models, the vector genome titer from the bioreactor was improved to beyond 1 × 1012 vg/mL. Five critical parameters were identified that had large effects on the pre-purification vector quantity—the transfection pH, production pH, complexation time, viable cell density at transfection, and transfection reagent to DNA ratio. The optimized process was further assessed for its performance extending to six AAV serotypes, namely AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9 carrying a transgene encoding for green fluorescent protein (GFP). Five of the six serotypes returned higher vector genome titers than the control condition. These data suggest that the choice of transfection reagent is a major factor in improving vector yield. The multivariate design of experiment approach is a powerful way to optimize production processes, and the optimized process from one AAV vector can to some extent be generalized to other serotypes and transgenes to accelerate development timelines of new programs.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3445","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140048611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Leveraging bioanalytical characterization of fractionated monoclonal antibody pools to identify aggregation-prone and less filterable proteoforms during virus filtration 利用分馏单克隆抗体池的生物分析特征，识别病毒过滤过程中易聚集和不易过滤的蛋白形式。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-03-07 DOI: 10.1002/btpr.3451

Solomon Isu, Lilia Vinskus, Derek Silva, Kristina Cunningham, Thomas Elich, Patricia Greenhalgh, Adam Sokolnicki, Bala Raghunath

{"title":"Leveraging bioanalytical characterization of fractionated monoclonal antibody pools to identify aggregation-prone and less filterable proteoforms during virus filtration","authors":"Solomon Isu, Lilia Vinskus, Derek Silva, Kristina Cunningham, Thomas Elich, Patricia Greenhalgh, Adam Sokolnicki, Bala Raghunath","doi":"10.1002/btpr.3451","DOIUrl":"10.1002/btpr.3451","url":null,"abstract":"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 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. Opportunities for molecular-level prediction of mAb filterability are needed to plug the existing knowledge gap in downstream processing. A molecular-level approach to understanding the factors influencing mAb filterability may reduce process development time, material loss, and processing costs due to oversized virus filters. In this work, pH step gradient fractionation was applied on polished bulk mAb feed to obtain concentrated pools of fractionated mAb variants. Biophysical properties and quality attributes of fractionated pools, including oligomeric state (size), isoelectric point profile, diffusion interaction parameters, and glycoform profile, were determined using bioanalytical methods. Filterability (loading and throughput) of fractionated pools were evaluated. Statistical methods were used to obtain correlations between quality attributes of mAb fractions and filterability on the Viresolve Pro virus filter.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3451","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140048668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Fed-batch performance profiles for mAb production using different intensified N − 1 seed strategies are CHO cell-line dependent 使用不同的强化 N - 1 种子策略生产 mAb 的联产批次性能曲线取决于 CHO 细胞系。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-28 DOI: 10.1002/btpr.3446

Yawen Tang, Jianlin Xu, Mengmeng Xu, Zhuangrong Huang, Johanna Santos, Qin He, Michael Borys, Anurag Khetan

{"title":"Fed-batch performance profiles for mAb production using different intensified N − 1 seed strategies are CHO cell-line dependent","authors":"Yawen Tang, Jianlin Xu, Mengmeng Xu, Zhuangrong Huang, Johanna Santos, Qin He, Michael Borys, Anurag Khetan","doi":"10.1002/btpr.3446","DOIUrl":"10.1002/btpr.3446","url":null,"abstract":"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 (enriched-batch or fed-batch) or perfusion culture to reach those higher final N − 1 viable cell densities (VCD). In this study, we evaluated how different N − 1 intensification strategies, specifically enriched-batch (EB) N − 1 versus perfusion N − 1, affect cell growth profiles and monoclonal antibody (mAb) productivity in the final N-stage production bioreactor operated in fed-batch mode. Three representative Chinese Hamster Ovary (CHO) cell lines producing different mAbs were cultured using either EB or perfusion N − 1 seeds and found that the N-stage cell growth and mAb productivities were comparable between EB N − 1 and perfusion N − 1 conditions for two of the cell lines but were very different for the third. In addition, within the two similar cell growth cell lines, differences in cell-specific productivity were observed. This suggests that the impact of the N − 1 intensification process on production was cell-line dependent. This study revealed that the N − 1 intensification strategy and the state of seeds from the different N − 1 conditions may affect the outcome of the N production stage, and thus, the choice of N − 1 intensification strategy could be a new target for future upstream optimization of mAb production.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"40 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139982288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0