Biotechnology Progress最新文献_第3页

Just passing through: Deploying aquaporins in microbial cell factories. 只是经过：在微生物细胞工厂中部署水汽蛋白

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-07-25 DOI: 10.1002/btpr.3497

Liam Richard Jenkins Sánchez, Lobke Maria Sips, Inge Noëlle Adriënne Van Bogaert

{"title":"Just passing through: Deploying aquaporins in microbial cell factories.","authors":"Liam Richard Jenkins Sánchez, Lobke Maria Sips, Inge Noëlle Adriënne Van Bogaert","doi":"10.1002/btpr.3497","DOIUrl":"https://doi.org/10.1002/btpr.3497","url":null,"abstract":"As microbial membranes are naturally impermeable to even the smallest biomolecules, transporter proteins are physiologically essential for normal cell functioning. This makes transporters a key target area for engineering enhanced cell factories. As part of the wider cellular transportome, aquaporins (AQPs) are responsible for transporting small polar solutes, encompassing many compounds which are of great interest for industrial biotechnology, including cell feedstocks, numerous commercially relevant polyols and even weak organic acids. In this review, examples of cell factory engineering by targeting AQPs are presented. These AQP modifications aid in redirecting carbon fluxes and boosting bioconversions either by enhanced feedstock uptake, improved intermediate retention, increasing product export into the media or superior cell viability against stressors with applications in both bacterial and yeast production platforms. Additionally, the future potential for AQP deployment and targeting is discussed, showcasing hurdles and considerations of this strategy as well as recent advances and future directions in the field. By leveraging the natural diversity of AQPs and breakthroughs in channel protein engineering, these transporters are poised to be promising tools capable of enhancing a wide variety of biotechnological processes.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3497"},"PeriodicalIF":2.5,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141756911","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

Identification of infectious viruses for risk-based virus testing of CHO unprocessed bulk using next-generation sequencing. 利用新一代测序技术识别传染性病毒，对未经加工的散装 CHO 进行基于风险的病毒检测。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-07-25 DOI: 10.1002/btpr.3485

Tiffany Hsu, Mary Jo Talley, Ping Yang, Angela Geiselhoeringer, Cindy Yang, Aditya Gorla, M Julhasur Rahman, Lindsey Silva, Dayue Chen, Bin Yang

{"title":"Identification of infectious viruses for risk-based virus testing of CHO unprocessed bulk using next-generation sequencing.","authors":"Tiffany Hsu, Mary Jo Talley, Ping Yang, Angela Geiselhoeringer, Cindy Yang, Aditya Gorla, M Julhasur Rahman, Lindsey Silva, Dayue Chen, Bin Yang","doi":"10.1002/btpr.3485","DOIUrl":"https://doi.org/10.1002/btpr.3485","url":null,"abstract":"It is important to increase manufacturing speed to make medicines more widely available. One bottleneck for CHO-based drug substance release is the in vitro viral (IVV) cell-based assay on unprocessed bulk. To increase process speed, we evaluate the suitability of replacing the IVV cell-based assay with next-generation sequencing (NGS). First, we outline how NGS is currently used in the pharmaceutical industry, and how it may apply to CHO virus testing. Second, we examine CHO virus contamination history. Since prior virus contaminants can replicate in the production bioreactor, we perform a literature search and classify 159 viruses as high, medium, low, or unknown risk based on their ability to infect CHO cells. Overall, the risk of virus contamination during the CHO manufacturing process is low. Only six viruses were reported to have contaminated CHO bioprocesses over the past several decades, and were primarily caused by fetal bovine serum or cell culture components. These virus contamination events can be mitigated through limitation and control of raw materials, combined with virus testing and virus clearance technologies. The list of CHO infectious viruses provides a starting framework for virus safety risk assessment and NGS development. Furthermore, ICH Q5A (R2) includes NGS as a molecular method for adventitious agent testing, paving a path forward for modernizing CHO virus testing.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3485"},"PeriodicalIF":2.5,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141756910","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 two transposases for improving expression of recombinant proteins in Chinese hamster ovary cell stable pools by co-transfection and supertransfection approaches. 评估两种转座酶通过共转染和超转染方法改善重组蛋白在中国仓鼠卵巢细胞稳定池中的表达。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-07-17 DOI: 10.1002/btpr.3496

Melina Lenser, Hanh Giai Ngo, Lily Sarrafha, Yashas Rajendra

{"title":"Evaluation of two transposases for improving expression of recombinant proteins in Chinese hamster ovary cell stable pools by co-transfection and supertransfection approaches.","authors":"Melina Lenser, Hanh Giai Ngo, Lily Sarrafha, Yashas Rajendra","doi":"10.1002/btpr.3496","DOIUrl":"https://doi.org/10.1002/btpr.3496","url":null,"abstract":"Transposons are genetic elements capable of cutting and pasting genes of interest via the action of a transposase and offer many advantages over random or targeted integration of DNA in the creation of Chinese hamster ovary (CHO) cell lines for recombinant protein expression. Unique transposases have different recognition sites, allowing multiple transposases to be co-transfected together. They also allow for supertransfection (transfection on a previously transfected pool or cell line) with a second transposase to integrate additional copies of the same gene or an additional gene without disruption of the previously integrated DNA which to our knowledge has not been previously described in literature. Two fluorescent proteins, EGFP and tagRFP657, were either co-transfected or supertransfected into CHO cells using two unique transposases and showed high expression efficiency with similar expression levels (measured as mean fluorescence intensity), regardless of whether the genes were co-transfected or supertransfected onto an existing stable pool. Additionally, dual selection of the genes, both in the absence of L-glutamine and the presence of puromycin, led to higher expression levels than single selection alone. These results demonstrate that supertransfection using unique transposases could be a useful strategy for increasing titers of existing cell lines or for overexpressing helper (non-therapeutic) genes to improve expression and/or product quality of existing pools and cell lines, potentially saving significant time and resources.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3496"},"PeriodicalIF":2.5,"publicationDate":"2024-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141625867","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

Proteomics-based method to comprehensively model the removal of host cell protein impurities. 基于蛋白质组学的方法，全面模拟清除宿主细胞蛋白质杂质的过程。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-07-17 DOI: 10.1002/btpr.3494

Roxana Disela, Daphne Keulen, Eleni Fotou, Tim Neijenhuis, Olivier Le Bussy, Geoffroy Geldhof, Martin Pabst, Marcel Ottens

{"title":"Proteomics-based method to comprehensively model the removal of host cell protein impurities.","authors":"Roxana Disela, Daphne Keulen, Eleni Fotou, Tim Neijenhuis, Olivier Le Bussy, Geoffroy Geldhof, Martin Pabst, Marcel Ottens","doi":"10.1002/btpr.3494","DOIUrl":"https://doi.org/10.1002/btpr.3494","url":null,"abstract":"Mechanistic models mostly focus on the target protein and some selected process- or product-related impurities. For a better process understanding, however, it is advantageous to describe also reoccurring host cell protein impurities. Within the purification of biopharmaceuticals, the binding of host cell proteins to a chromatographic resin is far from being described comprehensively. For a broader coverage of the binding characteristics, large-scale proteomic data and systems level knowledge on protein interactions are key. However, a method for determining binding parameters of the entire host cell proteome to selected chromatography resins is still lacking. In this work, we have developed a method to determine binding parameters of all detected individual host cell proteins in an Escherichia coli harvest sample from large-scale proteomics experiments. The developed method was demonstrated to model abundant and problematic proteins, which are crucial impurities to be removed. For these 15 proteins covering varying concentration ranges, the model predicts the independently measured retention time during the validation gradient well. Finally, we optimized the anion exchange chromatography capture step in silico using the determined isotherm parameters of the persistent host cell protein contaminants. From these results, strategies can be developed to separate abundant and problematic impurities from the target antigen.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3494"},"PeriodicalIF":2.5,"publicationDate":"2024-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141625868","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 comparison of different intensified upstream processes highlighting the advantage of WuXi Biologics' Ultra-high Productivity platform (WuXiUP^TM) in improved product quality and purification yield. 不同强化上游工艺的比较，凸显药明康德超高生产率平台（WuXiUPTM）在提高产品质量和纯化率方面的优势。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-07-09 DOI: 10.1002/btpr.3487

Xiang Zheng, Mingyue Fang, Yanling Zou, Shuo Wang, Weichang Zhou, Hang Zhou

{"title":"A comparison of different intensified upstream processes highlighting the advantage of WuXi Biologics' Ultra-high Productivity platform (WuXiUPTM) in improved product quality and purification yield.","authors":"Xiang Zheng, Mingyue Fang, Yanling Zou, Shuo Wang, Weichang Zhou, Hang Zhou","doi":"10.1002/btpr.3487","DOIUrl":"https://doi.org/10.1002/btpr.3487","url":null,"abstract":"WuXiUPTM, WuXi Biologics' Ultra-high Productivity platform, is an intensified and integrated continuous bioprocess platform developed for production of various biologics including monoclonal antibodies, fusion proteins, and bispecific antibodies. This process technology platform has manifested its remarkable capability in boosting the volumetric productivity of various biologics and has been implemented for large-scale clinical material productions. In this paper, case studies of the production of different pharmaceutical proteins using two high-producing and intensified culture modes of WuXiUPTM and the concentrated fed-batch (CFB), as well as the traditional fed-batch (TFB) are discussed from the perspectives of cell growth, productivity, and protein quality. Both WuXiUPTM and CFB outperformed TFB regarding volumetric productivity. Additionally, distinctive advantages in product quality profiles in the WuXiUPTM process, such as reduced acidic charge variants and fragmentation, are revealed. Therefore, a simplified downstream purification process with only two chromatographic steps can be developed to deliver the target product at a satisfactory purity and an extremely-high yield.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3487"},"PeriodicalIF":2.5,"publicationDate":"2024-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141557945","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

Rapid total sialic acid monitoring during cell culture process using a machine learning model based soft sensor. 利用基于机器学习模型的软传感器在细胞培养过程中快速监测总硅酸。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-07-02 DOI: 10.1002/btpr.3493

Amir M Behdani, Yuxiang Zhao, Grace Yao, Dhanuka Wasalathanthri, Eric Hodgman, Michael Borys, Gloria Li, Anurag Khetan, Dayanjan Wijesinghe, Anthony Leone

{"title":"Rapid total sialic acid monitoring during cell culture process using a machine learning model based soft sensor.","authors":"Amir M Behdani, Yuxiang Zhao, Grace Yao, Dhanuka Wasalathanthri, Eric Hodgman, Michael Borys, Gloria Li, Anurag Khetan, Dayanjan Wijesinghe, Anthony Leone","doi":"10.1002/btpr.3493","DOIUrl":"https://doi.org/10.1002/btpr.3493","url":null,"abstract":"Total sialic acid content (TSA) in biotherapeutic proteins is often a critical quality attribute as it impacts the drug efficacy. Traditional wet chemical assays to quantify TSA in biotherapeutic proteins during cell culture typically takes several hours or longer due to the complexity of the assay which involves isolation of sialic acid from the protein of interest, followed by sample preparation and chromatographic based separation for analysis. Here, we developed a machine learning model-based technology to rapidly predict TSA during cell culture by using typically measured process parameters. The technology features a user interface, where the users only have to upload cell culture process parameters as input variables and TSA values are instantly displayed on a dashboard platform based on the model predictions. In this study, multiple machine learning algorithms were assessed on our dataset, with the Random Forest model being identified as the most promising model. Feature importance analysis from the Random Forest model revealed that attributes like viable cell density (VCD), glutamate, ammonium, phosphate, and basal medium type are critical for predictions. Notably, while the model demonstrated strong predictability by Day 14 of observation, challenges remain in forecasting TSA values at the edges of the calibration range. This research not only emphasizes the transformative power of machine learning and soft sensors in bioprocessing but also introduces a rapid and efficient tool for sialic acid prediction, signaling significant advancements in bioprocessing. Future endeavors may focus on data augmentation to further enhance model precision and exploration of process control capabilities.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3493"},"PeriodicalIF":2.5,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141475819","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

Biotechnological potential of yeast cell wall: An overview. 酵母细胞壁的生物技术潜力：综述。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-06-27 DOI: 10.1002/btpr.3491

Fanny Machado Jofre, Sarah de Souza Queiroz, Diana Alva Sanchez, Priscila Vaz Arruda, Júlio César Dos Santos, Maria das Graças de Almeida Felipe

{"title":"Biotechnological potential of yeast cell wall: An overview.","authors":"Fanny Machado Jofre, Sarah de Souza Queiroz, Diana Alva Sanchez, Priscila Vaz Arruda, Júlio César Dos Santos, Maria das Graças de Almeida Felipe","doi":"10.1002/btpr.3491","DOIUrl":"https://doi.org/10.1002/btpr.3491","url":null,"abstract":"The yeast cell wall is a complex structure whose main function is to protect the cell from physical and chemical damage, providing it with rigidity. It is composed of a matrix of covalently linked polysaccharides and proteins, including β-glucans, mannoproteins, and chitin, whose proportion can vary according to the yeast species and environmental conditions. The main components of the yeast cell wall have relevant properties that expand the possibilities of use in different industrial sectors, such as pharmaceutical, food, medical, veterinary, and cosmetic. Some applications include bioremediation, enzyme immobilization, animal feed, wine production, and hydrogel production. In the literature it is the description of the cell wall composition of model species like Saccharomyces cerevisiae and Candida albicans, however, it is important to know that this composition can vary according to the species or the culture medium conditions. Thus, understanding the structural composition of different species holds promise as an alternative to expanding the utilization of residual yeast from different bioprocesses. In the context of a circular economy, the conversion of residual yeast into valuable products is an attractive prospect for researchers aiming to develop sustainable technologies. This review provides an overview of yeast cell wall composition and its significance in biotechnological applications, considering prospects to increase the diversification of these compounds in industry.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3491"},"PeriodicalIF":2.5,"publicationDate":"2024-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141454908","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

Modeling of cell cultivation for monoclonal antibody production processes considering lactate metabolic shifts. 考虑乳酸代谢转变的单克隆抗体生产过程的细胞培养模型。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-06-24 DOI: 10.1002/btpr.3486

Kozue Okamura, Sara Badr, Yusuke Ichida, Akira Yamada, Hirokazu Sugiyama

{"title":"Modeling of cell cultivation for monoclonal antibody production processes considering lactate metabolic shifts.","authors":"Kozue Okamura, Sara Badr, Yusuke Ichida, Akira Yamada, Hirokazu Sugiyama","doi":"10.1002/btpr.3486","DOIUrl":"https://doi.org/10.1002/btpr.3486","url":null,"abstract":"Demand for monoclonal antibodies (mAbs) is rapidly increasing. To achieve higher productivity, there have been improvements to cell lines, operating modes, media, and cultivation conditions. Representative mathematical models are needed to narrow down the growing number of process alternatives. Previous studies have proposed mechanistic models to depict cell metabolic shifts (e.g., lactate production to consumption). However, the impacts of variations of some operating conditions have not yet been fully incorporated in such models. This paper offers a new mechanistic model considering variations in dissolved oxygen and glutamine depletion on cell metabolism applied to a novel Chinese hamster ovary (CHO) cell line. Expressions for the specific rates of lactate production, glutamine consumption, and mAb production were formulated for stirred and shaken-tank reactors. A deeper understanding of lactate metabolic shifts was obtained under different combinations of experimental conditions. Lactate consumption was more pronounced in conditions with higher DO and low glutamine concentrations. The model offers mechanistic insights that are useful for designing advanced operation strategies. It can be used in design space generation and process optimization for better productivity and product quality.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3486"},"PeriodicalIF":2.5,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141454909","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

Cover 封面

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-06-24 DOI: 10.1002/btpr.3488

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

引用次数: 0

Primary clarification of mammalian cell culture fluid using enhanced sedimentation on inclined surfaces inside the single-use disposable Sudhin BioSettler150. 利用一次性使用的 Sudhin BioSettler150 内倾斜表面上的增强沉降功能，对哺乳动物细胞培养液进行初级澄清。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-06-19 DOI: 10.1002/btpr.3489

Christopher B Glascock, Dhinakar S Kompala

{"title":"Primary clarification of mammalian cell culture fluid using enhanced sedimentation on inclined surfaces inside the single-use disposable Sudhin BioSettler150.","authors":"Christopher B Glascock, Dhinakar S Kompala","doi":"10.1002/btpr.3489","DOIUrl":"10.1002/btpr.3489","url":null,"abstract":"The first downstream processing step in the purification of a biopharmaceutical protein secreted into mammalian cell culture fluid is the primary clarification of the culture fluid. As cell densities in the fed-batch and increasingly more common perfusion bioreactors have increased over last two decades through intensified upstream bioreactor production processes, the traditional primary clarification unit operations of centrifugation and/or microfiltration become more challenging with issues like frequent desludging, cell disruption due to shear damage and quick fouling of membranes. We have developed a novel compact cell settler device exploiting the enhanced sedimentation on inclined surfaces and demonstrated that this settler device can be adapted easily to remove and contain cells or cell clumps from the clarified supernatant collected via the top effluent of the settler. In this work, we present high product recovery results during primary clarification of mammalian cell culture supernatant using our novel single-use disposable BioSettler150 while processing about 10 L of cell culture broth within short processing times of about 4 h.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e3489"},"PeriodicalIF":2.5,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141426249","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