Biotechnology Progress最新文献_第9页

Effect of inner diameter, filter length, and pore size on hollow fiber filter fouling during perfusion cell culture 内径、过滤器长度和孔径对灌流细胞培养过程中中空纤维过滤器堵塞的影响。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-11 DOI: 10.1002/btpr.3440

Dominique WuDunn, Andrea Squeri, Jimmy Vu, Ashna Dhingra, Jon Coffman, Ken Lee

{"title":"Effect of inner diameter, filter length, and pore size on hollow fiber filter fouling during perfusion cell culture","authors":"Dominique WuDunn, Andrea Squeri, Jimmy Vu, Ashna Dhingra, Jon Coffman, Ken Lee","doi":"10.1002/btpr.3440","DOIUrl":"10.1002/btpr.3440","url":null,"abstract":"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) past process limitations. In this work, the impact of hollow fiber filter geometries on product sieving and hydraulic membrane resistance profiles is evaluated in a tangential flow filtration (TFF) perfusion system. The hollow fibers tested had lengths ranging from 19.8 to 41.5 cm, inner diameters (IDs) ranging from 1.0 to 2.6 mm, and pore sizes of 0.2 or 0.65 μm. The results showed that the shortest hollow fibers experienced higher product sieving while larger IDs contributed to both higher product sieving and lower hydraulic membrane resistances, illustrating the impact of filter geometry on process performance. The results also showed 0.2 μm pore size filters maintain higher product sieving, but also higher membrane resistances compared to 0.65 μm pore size filters. This study highlights the need for optimized hollow fiber filter geometries to maximize use of the membrane area, which in turn can reduce production costs and increase scalability of the perfusion process.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3440","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139721471","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

Advancing multiproduct resin reuse for development and clinical manufacturing of an antibody-based therapeutic 推进多产品树脂再利用，用于抗体疗法的开发和临床生产。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-09 DOI: 10.1002/btpr.3434

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

{"title":"Advancing multiproduct resin reuse for development and clinical manufacturing of an antibody-based therapeutic","authors":"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","doi":"10.1002/btpr.3434","DOIUrl":"10.1002/btpr.3434","url":null,"abstract":"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 such as the COVID-19 pandemic. The work presented herein describes an overarching multiproduct resin reuse (MRR) strategy, which includes a risk assessment, strategic planning, small-scale feasibility runs, and the successful execution of the MRR strategy to support Good manufacturing practice (GMP) clinical manufacturing of an antibody-based therapeutic. Specifically, an anion exchange (AEX) and cation exchange (CEX) MRR strategy is described. Clearance of carryover biological product is demonstrated by first cleaning the AEX and CEX manufacturing columns with sodium hydroxide to ensure inactivation and degradation of the carryover protein and followed by a blank buffer elution that is tested using various analytical methodologies to ensure reduction of the carryover protein to an acceptable level. To our knowledge, this is the first time an MRR approach has been successfully implemented and submitted to health authorities to support biologic GMP clinical manufacture.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3434","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139705918","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

Implementation of mDoE-methods to a microcarrier-based expansion processes for mesenchymal stem cells 将 mDoE 方法应用于基于微载体的间充质干细胞扩增过程。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-09 DOI: 10.1002/btpr.3429

Kim B. Kuchemüller, Ralf Pörtner, Johannes Möller

{"title":"Implementation of mDoE-methods to a microcarrier-based expansion processes for mesenchymal stem cells","authors":"Kim B. Kuchemüller, Ralf Pörtner, Johannes Möller","doi":"10.1002/btpr.3429","DOIUrl":"10.1002/btpr.3429","url":null,"abstract":"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 of microcarrier-based expansion systems. One accelerating approach for process design is model-assisted Design of Experiments (mDoE) combining mathematical process models and statistical tools. In this study, the mDoE workflow was used for the development of an expansion processes with human immortalized mesenchymal stem cells (hMSC-TERT) and the aim of maximizing cell yield assuming only a limited amount of prior knowledge at a very early stage of development. First, suitable microcarriers for expansion in shake flasks were screened and the differentiation of the cells was proven. Second, initial experiments were performed to generate prior knowledge, which was then used to set up the mathematical model and to estimate the model parameters. Finally, the mDoE was used to determine and evaluate the design space to be performed experimentally. Overall, a cell expansion process using microcarriers in a shake flask culture was successfully implemented and a significant increase in cell yield (up to 6,2-fold) was achieved compared to literature.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3429","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139705919","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

Effect of flux and shear rate on E. coli recovery in tangential flow filtration through a single hollow fiber 流量和剪切率对单根中空纤维切向流过滤中大肠杆菌回收率的影响。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-08 DOI: 10.1002/btpr.3432

Jessica Zuponcic, Fernanda Cunha, Grant Springer, Eduardo Ximenes, Michael R. Ladisch

{"title":"Effect of flux and shear rate on E. coli recovery in tangential flow filtration through a single hollow fiber","authors":"Jessica Zuponcic, Fernanda Cunha, Grant Springer, Eduardo Ximenes, Michael R. Ladisch","doi":"10.1002/btpr.3432","DOIUrl":"10.1002/btpr.3432","url":null,"abstract":"Pathogenic bacteria which enter a viable but non-culturable (VBNC) state impede efforts to reach detectable concentrations required for PCR methods. This motivated a strategy for tangential flow filtration to concentrate bacteria in aqueous samples while maintaining the bacteria in a viable state, maximizing their recovery and achieving high fluxes through a single hollow fiber membrane. Filtrations were carried out for green fluorescent protein (GFP) E. coli at high shear rates (up to 27,000 sec−1) through 0.2 μm cut-off polyethersulfone (PES) microfilter membranes or 50 kDa polysulfone (PS) ultrafilter membranes. High shear minimized bacterial attachment on membrane surfaces, which would otherwise occur due to forced convection of the particles to the membrane surface at high flux conditions. Single fiber filter modules were constructed to facilitate concentration of Escherichia coli at fluxes ranging from 55 to 4500 L m−2 h−1. The effect of high shear rates on bacterial viability was found to be minimal with bacterial losses during filtration caused principally by their accumulation on the membrane surface. Recoveries of 90% were achievable at high shear rates when the average flux was ≤300 L m−2 h−1. This corresponded to a 3-h filtration time for a 225 mL sample through a single hollow fiber. Detectable bacteria concentrations of 1800 colony-forming unit (CFU)/mL were achieved for starting concentrations of 140 CFU/mL.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3432","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139701695","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

Demonstration of a robust high cell density transient CHO platform yielding mAb titers of up to 2 g/L without medium exchange 展示了一个强大的高细胞密度瞬时 CHO 平台，该平台无需更换培养基即可获得高达 2 克/升的 mAb 滴度。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-08 DOI: 10.1002/btpr.3435

Rigumula Wu, Danielle M. Kahl, Ronald Kloberdanz, Kushal J. Rohilla, Sowmya Balasubramanian

{"title":"Demonstration of a robust high cell density transient CHO platform yielding mAb titers of up to 2 g/L without medium exchange","authors":"Rigumula Wu, Danielle M. Kahl, Ronald Kloberdanz, Kushal J. Rohilla, Sowmya Balasubramanian","doi":"10.1002/btpr.3435","DOIUrl":"10.1002/btpr.3435","url":null,"abstract":"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 Chinese hamster ovary (CHO) is crucial. It maintains the consistency in cell lines and processes throughout the early drug discovery and downstream manufacturing processes. This helps researchers to identify the issues at an early stage for timely resolution. In this study, we have demonstrated a simple high-titer platform for TGE in CHO based on a dilution process of seeding cells. We achieved titers ranging from 0.8 to 1.9 g/L for eight model mAbs at three scales (1, 30, 100 mL) in 10 days using our new platform. The ability to seed by dilution significantly streamlined the process and dramatically enhanced platform throughput. We observed a modest reduction in titer ranging from 11% to 28% when cells were seeded using dilution compared to when cells were seeded using medium exchange. Further studies revealed that carry over of spent medium into transfection negatively affected the DNA uptake and transcription processes, while the translation and secretion was minimally impacted. In summary, our transient CHO platform using cells prepared by dilution at high densities can achieve high titers of up to 1.9 g/L, which can be further improved by targeting the bottlenecks of transfection and transcription.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139701694","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

Gene copy number, gene configuration and LC/HC mRNA ratio impact on antibody productivity and product quality in targeted integration CHO cell lines 基因拷贝数、基因配置和 LC/HC mRNA 比率对靶向整合 CHO 细胞系抗体生产率和产品质量的影响。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-02-06 DOI: 10.1002/btpr.3433

Zion Lee, Jun Wan, Amy Shen, Gavin Barnard

{"title":"Gene copy number, gene configuration and LC/HC mRNA ratio impact on antibody productivity and product quality in targeted integration CHO cell lines","authors":"Zion Lee, Jun Wan, Amy Shen, Gavin Barnard","doi":"10.1002/btpr.3433","DOIUrl":"10.1002/btpr.3433","url":null,"abstract":"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 of both IgG heavy chain (HC) and light chain (LC) concurrently resulted in the attainment of plateaus in mRNA levels and product titers, notably occurring beyond four to five gene copies integrated at the same TI site. Furthermore, maintaining a fixed gene copy number while varying the position of genes within the vector influenced the LC/HC mRNA ratio, which subsequently exerted a substantial impact on product titer. Moreover, manipulation of the LC/HC gene ratio through the introduction of surplus LC gene copies led to heightened LC mRNA expression and a reduction in the levels of high molecular weight species. It is noteworthy that the effects of excess LC on product titer were dependent on the specific molecule under consideration. The strategic utilization of PCR tags enabled precise quantification of transcription from each expression slot within the vector, facilitating the identification of highly expressive and less expressive slots. Collectively, these findings significantly enhance our understanding of stable antibody production in TI CHO cell lines.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139696909","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

Correction to “Investigation of the hepatic respiration and liver zonation on rat hepatocytes using an integrated oxygen biosensor in a microscale device” 对 "利用微型装置中的集成氧生物传感器研究大鼠肝细胞的肝呼吸和肝分区 "的更正。

IF 2.9 3区生物学

Biotechnology Progress Pub Date : 2024-01-30 DOI: 10.1002/btpr.3422

引用次数: 0

Development of bioreactor scale-down model using orthogonal projections to latent structures method and CO2 supplementation 利用正交投影潜伏结构法和二氧化碳补充法开发生物反应器缩小模型。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-01-30 DOI: 10.1002/btpr.3423

Jinxin Gao, Laurie B. Hazeltine, Neal Stroud, Ning Liu, Yao-ming Huang

{"title":"Development of bioreactor scale-down model using orthogonal projections to latent structures method and CO2 supplementation","authors":"Jinxin Gao, Laurie B. Hazeltine, Neal Stroud, Ning Liu, Yao-ming Huang","doi":"10.1002/btpr.3423","DOIUrl":"10.1002/btpr.3423","url":null,"abstract":"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 method, orthogonal projections to latent structures (OPLS) analysis, can be utilized to compare time-course cell culture data across scales. Here we describe an example where OPLS is used to identify gaps between small-scale and large-scale bioreactor performances. In this case, differences in the partial pressure of carbon dioxide (pCO2) and lactate profiles were observed between small- and large-scale bioreactors, which were linked to differences in the product-quality attributes fragments and galactosylation. An improved small-scale model was developed, leading to improved consistency in the process performance and product qualities across scales and qualification of the scale-down model for regulatory submissions. This new statistical approach can provide valuable insights into process understanding and process scale-up.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139575177","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

Process development and characterization for integrated continuous bioprocesses—Highlights from N-mAb 集成连续生物工艺的过程开发和表征--来自 N-mAb 的亮点。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-01-30 DOI: 10.1002/btpr.3425

Kevin Brower, Kelly Wiltberger, Claudia Berdugo, Allen Bosley, Elizabeth Goodrich, Valerie Pferdeort, Gene Schaefer

引用次数: 0

Enhancing cell separation in a hybrid spiral dielectrophoretic microchannel: Numerical insights and optimal operating conditions 加强混合螺旋介电泳微通道中的细胞分离：数值分析和最佳操作条件。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2024-01-30 DOI: 10.1002/btpr.3437

Mohammed Raihan Uddin, Xiaolin Chen

{"title":"Enhancing cell separation in a hybrid spiral dielectrophoretic microchannel: Numerical insights and optimal operating conditions","authors":"Mohammed Raihan Uddin, Xiaolin Chen","doi":"10.1002/btpr.3437","DOIUrl":"10.1002/btpr.3437","url":null,"abstract":"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 work for continuous and single-stage separation of lung cancer cell line A549 cells from white blood cells of overlapping size. The working mechanism of the proposed spiral microchannel embedded with planar interdigitated electrodes is validated against the experimental results. A numerical investigation is carried out over a range of flow conditions and electric field intensity to determine the separation efficiency and migration characteristics of the cell mixture. The results demonstrate the unique capability of the proposed microchannel to achieve high-throughput separation of cells at low applied voltages in both vertical and lateral directions. A significant lateral separation distance between the CTCs and the WBCs has been achieved, which allows for high-resolution and effective separation of cells. The separation resolution can be controlled by adjusting the strength of the applied electric field. Furthermore, the results demonstrate that the lateral separation distance is maximum at a voltage termed the critical voltage, which increases with the increase in the flow rate. The proposed microchannel and the developed technique can provide valuable insight into the development of a tunable and robust medical device for effective and high-throughput separation of cancer cells from the WBCs.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btpr.3437","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139641553","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