Biotechnology Progress最新文献_第8页

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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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

Catechins prevent monoclonal antibody fragmentation during production via fed-batch culture of Chinese hamster ovary cells 儿茶素可防止中国仓鼠卵巢细胞批量喂养生产过程中出现单克隆抗体碎片。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-28 DOI: 10.1002/btpr.3447

Tsuyoshi Yamaguchi, Hiroko Ishikawa, Mie Fukuda, Yumi Sugita, Misaki Furuie, Ryuma Nagano, Toshiyuki Suzawa, Koichi Yamamoto, Kaori Wakamatsu

{"title":"Catechins prevent monoclonal antibody fragmentation during production via fed-batch culture of Chinese hamster ovary cells","authors":"Tsuyoshi Yamaguchi, Hiroko Ishikawa, Mie Fukuda, Yumi Sugita, Misaki Furuie, Ryuma Nagano, Toshiyuki Suzawa, Koichi Yamamoto, Kaori Wakamatsu","doi":"10.1002/btpr.3447","DOIUrl":"10.1002/btpr.3447","url":null,"abstract":"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 this study, we observed that the antibody fragment content increased with increasing titers in fed-batch cultures for all three cell lines expressing recombinant antibodies. Adding copper sulfate to the culture medium further increased the fragment content, suggesting the involvement of reactive oxygen species (ROS) in the fragmentation process. Though antioxidants may be helpful to scavenge ROS, several antioxidants are reported to decrease the productivity of CHO cells. Among the antioxidants examined, we observed that the addition of catechin or (−)-epigallocatechin gallate to the culture medium prevented fragmentation content by about 20% and increased viable cell density and titer by 30% and 10%, respectively. Thus, the addition of catechins or compounds of equivalent function would be beneficial for manufacturing therapeutic mAbs with a balance between high titers and good quality.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3447","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139982287","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

Application of fucosylation inhibitors for production of afucosylated antibody 应用岩藻糖基化抑制剂生产岩藻糖基化抗体。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-28 DOI: 10.1002/btpr.3438

Ping Xu, Yu Chuan Ou, Michael Smith, Jim Paulson, Michael A. Schmidt, Lakshmi Kandari, Rodney Parsons, Anurag Khetan

引用次数: 0

Biotransformation of maize bran-derived ferulic acid to vanillin using an adapted strain of Amycolatopsis sp. ATCC 39116 利用一株适应的 Amycolatopsis sp. ATCC 39116，将玉米麸皮衍生的阿魏酸生物转化为香兰素。

IF 2.9 3区生物学

Biotechnology Progress Pub Date : 2024-02-28 DOI: 10.1002/btpr.3417

Rasika V. Tupe, Nitesh K. Singh, Annamma A. Odaneth

{"title":"Biotransformation of maize bran-derived ferulic acid to vanillin using an adapted strain of Amycolatopsis sp. ATCC 39116","authors":"Rasika V. Tupe, Nitesh K. Singh, Annamma A. Odaneth","doi":"10.1002/btpr.3417","DOIUrl":"10.1002/btpr.3417","url":null,"abstract":"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 were evaluated for biotransformation to vanillin, employing Amycolatopsis sp. as a whole-cell biocatalyst. Initial adaptation studies were critical in improving ferulic acid assimilation and its conversion to vanillin by 65% and 56%, respectively by the fourth adaptation cycle. The effect of cell's physiological states and vanillic acid supplementation on vanillin production was studied using standard ferulic acid as a substrate in an effort to achieve further improvement in vanillin yield. In the presence of vanillic acid, 18 h cultured cells using 2 g/L of standard and isolated ferulic acid produced vanillin concentrations of up to 0.71 and 0.48 g/L, respectively. Furthermore, intermediates involved in the ferulic acid catabolic pathway and their interrelations were studied using GC–MS analysis. Results indicated that two different routes were involved in the catabolism of standard ferulic acid, and similar metabolic routes were observed for an isolated ferulic acid stream. These findings effectively evaluated isolated ferulic acid for sustainable vanillin production while reducing agro-industrial waste pollution.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139982286","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

A deep-well plate enabled automated high-throughput cell line development platform 深孔板自动化高通量细胞系开发平台。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-20 DOI: 10.1002/btpr.3442

Xiaoyan Tang, Jorge Quiroz, Yixiao Zhang, Jessica Pan, Zhong Lai, Zhimei Du, Ren Liu

{"title":"A deep-well plate enabled automated high-throughput cell line development platform","authors":"Xiaoyan Tang, Jorge Quiroz, Yixiao Zhang, Jessica Pan, Zhong Lai, Zhimei Du, Ren Liu","doi":"10.1002/btpr.3442","DOIUrl":"10.1002/btpr.3442","url":null,"abstract":"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 resort to screening a large number of cell pools and clones to increase the probability of identifying the ideal production cell line, which is a very laborious and resource-demanding process. Here we have developed a deep-well plate (DWP) enabled high throughput (DEHT) CLD platform using 24-well DWP (24DWP), liquid handler, and other automation components. This platform has capabilities covering the key steps of CLD including cell passaging, clone imaging and expansion, and fed-batch production. We are the first to demonstrate the suitability of 24DWP for CLD by confirming minimal well-to-well and plate-to-plate variability and the absence of well-to-well cross contamination. We also demonstrated that growth, production, and product quality of 24DWP cultures were comparable to those of conventional shake flask cultures. The DEHT platform enables scientists to screen five times more cultures than the conventional CLD platform, thus significantly decreases the resources needed to identify an ideal production cell line for biologics manufacturing.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139911925","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

Evaluation of PDL1 positive cancer cell-specific binding activity of recombinant anti-PDL1 scFv 评估重组抗 PDL1 scFv 的 PDL1 阳性癌细胞特异性结合活性。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-20 DOI: 10.1002/btpr.3439

Sun-Hee Kim, Hae-Min Park, Hee-Jin Jeong

引用次数: 0

Leveraging machine learning to dissect role of combinations of amino acids in modulating the effect of zinc on mammalian cell growth 利用机器学习剖析氨基酸组合在调节锌对哺乳动物细胞生长的影响中的作用。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-15 DOI: 10.1002/btpr.3436

Ujjiti Pandey, Indrani Madhugiri, Chetan Gadgil, Mugdha Gadgil

{"title":"Leveraging machine learning to dissect role of combinations of amino acids in modulating the effect of zinc on mammalian cell growth","authors":"Ujjiti Pandey, Indrani Madhugiri, Chetan Gadgil, Mugdha Gadgil","doi":"10.1002/btpr.3436","DOIUrl":"10.1002/btpr.3436","url":null,"abstract":"Although the contributions of individual components of cell culture media are largely known, their combinatorial effects are far less understood. Experiments varying one component at a time cannot identify combinatorial effects, and analysis of the large number of experiments required to decipher such effects is challenging. Machine learning algorithms can help in the analysis of such datasets to identify multi-component interactions. Zinc toxicity in vitro is known to change depending on amino acid concentration in the extracellular medium. Multiple amino acids are known to be involved in this protection. Thirty-two amino acid compositions were formulated to evaluate their effect on the growth of CHO cells under high zinc conditions. A sequential machine learning analysis methodology was used, which led to the identification of a set of amino acids (threonine, proline, glutamate, aspartate, asparagine, and tryptophan) contributing to protection from zinc. Our results suggest that a decrease in availability of these set of amino acids due to consumption may affect cell growth in media formulated with high zinc concentrations, and in contrast, normal levels of these amino acids are associated with better tolerance to high zinc concentration. Our sequential analysis method may be similarly employed for high throughput medium design and optimization experiments to identify interactions among a large number of cell culture medium components.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139734328","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