Biotechnology Progress最新文献_第9页

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-04-07

引用次数: 0

Single-cell RNA sequencing reveals population-based shifts in gene expression heterogeneity in an unstable, clonally derived CHO cell line. 单细胞RNA测序揭示了不稳定的克隆来源的CHO细胞系中基因表达异质性的群体变化。

IF 2.8 3区生物学

Biotechnology Progress Pub Date : 2026-03-01 Epub Date: 2026-03-09 DOI: 10.1002/btpr.88502

Theodore W Peters, Jake T Leinas, Sofie O'Brien, Helena Ochoa-Moreno, Daniel Lachance, Patrick Younan, Greg Szeto, Howard Clarke

{"title":"Single-cell RNA sequencing reveals population-based shifts in gene expression heterogeneity in an unstable, clonally derived CHO cell line.","authors":"Theodore W Peters, Jake T Leinas, Sofie O'Brien, Helena Ochoa-Moreno, Daniel Lachance, Patrick Younan, Greg Szeto, Howard Clarke","doi":"10.1002/btpr.88502","DOIUrl":"10.1002/btpr.88502","url":null,"abstract":"The genomic plasticity of Chinese Hamster Ovary cells allows them to be genetically engineered to produce foreign proteins with high specific productivities while placing a significant burden on their ability to retain this expression capacity over time. Many cell lines lose expression over time and need to be screened for expression consistency. Stability has been studied in clonally derived cell cultures by evaluating phenotypic heterogeneity in subclones derived from stable or unstable clones, but not well characterized within the primary, clonally derived cell culture. To address this, we employed single cell RNA sequencing in stable and unstable cell lines expressing the same monoclonal antibody, at early and late timepoints in their manufacturing lifespan. We observed higher expression heterogeneity and a more drastic redistribution of expression profile \"clusters\" over serial passaging in the unstable cell line. Differentially expressed genes in those clusters making up the unstable cell line suggest distinct changes in metabolic pathway activity in this cell line over time. Using molecular methods, changing gene expression profiles were confirmed and activation of molecular pathways consistent with a challenged protein homeostasis were shown to be enriched in cells over time in the unstable cell line. This report more broadly implicates the role of heterogeneity within a clonal population as a contributing factor to cell line instability, providing a biological context for further investigation.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":" ","pages":"e88502"},"PeriodicalIF":2.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147389133","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

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-25

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IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-22

引用次数: 0

Exploring the design space for Triton X-100 substitutes in viral inactivation applications 探索Triton X-100替代品在病毒灭活应用中的设计空间。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-16 Epub Date: 2025-09-03 DOI: 10.1002/btpr.70069

Yuqi Du, Shanshan Wu

{"title":"Exploring the design space for Triton X-100 substitutes in viral inactivation applications","authors":"Yuqi Du, Shanshan Wu","doi":"10.1002/btpr.70069","DOIUrl":"10.1002/btpr.70069","url":null,"abstract":"The urgent need to replace the European-prohibited Triton X-100 in biomanufacturing has been hindered by insufficient data on alternative detergents' minimum effective concentrations (MECs) and process robustness in viral inactivation. This study makes systematic research including: (1) Establishment of MECs for novel Triton X-100 substitutes (TXR-1/VIS/13-S9/C16) achieving effective inactivation of Xenotropic murine leukemia virus and Pseudorabies virus (log10 reduction factor >4) across diverse CHO harvest fluids; (2) Demonstration of broad-spectrum efficacy against various viruses, with TXR-1/VIS/13-S9 maintaining effective inactivation for Bovine viral diarrhea virus, Vesicular stomatitis virus, Baculovirus, and Herpes simplex virus type 1; (3) Identification of PS20's material-dependent inactivation dynamics, establishing standalone parameters (4 h at 37°C) that achieve equivalent viral inactivation to traditional tri(n-butyl)phosphate -combined methods without requiring lipase activity—a paradigm shift in detergent application. Crucially, process optimization revealed that extending exposure time (1–4 h) enhanced PS20/PS80 efficacy more effectively than two fold concentration increases, providing cost-effective solutions. These findings deliver broader design spaces for implementing eco-friendly detergents while ensuring compliance with EMA/ICH viral safety standards.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 1","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940714","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

Enhanced deoxyviolacein bioproduction via enzyme condensation with a yeast glycolytic enzyme-derived peptide tag 通过酵母菌糖酵解酶衍生肽标签的酶缩合增强脱氧紫罗兰素的生物生产。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-16 Epub Date: 2025-10-11 DOI: 10.1002/btpr.70079

Rioko Ito, Sayoko Ito-Harashima, Naoko Segawa, Natsuko Miura, Michihiko Kataoka

{"title":"Enhanced deoxyviolacein bioproduction via enzyme condensation with a yeast glycolytic enzyme-derived peptide tag","authors":"Rioko Ito, Sayoko Ito-Harashima, Naoko Segawa, Natsuko Miura, Michihiko Kataoka","doi":"10.1002/btpr.70079","DOIUrl":"10.1002/btpr.70079","url":null,"abstract":"In microbial production, regulating endogenous and exogenous synthetic pathways is essential. Techniques to induce intracellular enzyme condensation have attracted attention as a means to increase apparent enzyme activity with minimal transcriptional and translational burden on the cell. Artificial enzyme condensation can be induced by tagging enzymes with specific proteins or peptides. In our previous study, we identified novel peptide tags derived from condensate-forming Saccharomyces cerevisiae glycolytic enzymes and their potential use in controlling intracellular metabolism by inducing artificial condensates in S. cerevisiae cells. Herein, we evaluated the condensate formation of two enzymes using peptide tags and the branched violacein biosynthetic pathway from Pseudoalteromonas luteoviolacea in S. cerevisiae, and tested the effects of modulating biosynthesis to increase the production of deoxyviolacein, a byproduct with antibacterial and anticancer activities. We used several protein and peptide tags with a simplified expression system, and all the tags successfully induced artificial condensate formation in the cell. Additionally, introducing a short peptide tag successfully increased deoxyviolacein production by approximately twofold, displaying a higher efficacy compared to FUSN, a previously reported N-terminal 213 amino acid region with an intrinsically disordered property. These results demonstrate the potential use of peptide tags to enhance bioproduction through the regulation of endogenous and exogenous synthetic pathways. The methods contribute to the development of novel strategies for microorganisms to be used for bioproduction through the controlled condensation of metabolic enzymes in cells.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 1","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12908088/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145273629","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

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-16

引用次数: 0

Evaluation of single-use disk stack continuous centrifuge to harvest monoclonal antibody from cell culture fluid 一次性圆盘叠置连续离心机从细胞培养液中获取单克隆抗体的评价。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-16 Epub Date: 2025-10-23 DOI: 10.1002/btpr.70084

Ailan Xu, Lei Gong, Chenglong Deng, Wangjian Sheng, Chen Hua, Bingxin Lu, Chen Li, Jun Ma, Jingbo Zhou, Xiao Zhang, Yiqing Cui

{"title":"Evaluation of single-use disk stack continuous centrifuge to harvest monoclonal antibody from cell culture fluid","authors":"Ailan Xu, Lei Gong, Chenglong Deng, Wangjian Sheng, Chen Hua, Bingxin Lu, Chen Li, Jun Ma, Jingbo Zhou, Xiao Zhang, Yiqing Cui","doi":"10.1002/btpr.70084","DOIUrl":"10.1002/btpr.70084","url":null,"abstract":"Various technologies, including precipitation, flocculation, depth filtration, microfiltration, and centrifugation, have been developed to clarify mammalian cell culture fluids. For processing volumes between 2000 and 5000 L, continuous centrifugation followed by depth filtration is the preferred method. This process starts with the removal of cells and large debris through continuous centrifugation, followed by the filtration of small debris and some impurities. The newly introduced single-use centrifuge, designed to prevent cross-contamination and mimic traditional continuous centrifuges, was evaluated for its performance, particularly focusing on its impact on cell lysis and subsequent filtration and purification processes. The single-use centrifuge showed better performance in reducing turbidity and lactate dehydrogenase levels (LDH) in the supernatant, indicating less cell lysis compared to the conventional centrifuge. A separation load factor range of 0.91–2.73 was identified as optimal for balancing centrifugation throughput and product quality. Both centrifuge types had a comparable impact on the performance of subsequent depth filtration, supporting a load capacity of at least 100 L/m2. No significant differences in product quality, including SE-HPLC, NR/R CE-SDS, icIEF, HCP, and rDNA, were observed between the conventional and single-use centrifuges. These harvest strategies did not affect the subsequent purification steps. For volumes up to 5000 L, both centrifuge types are viable; however, for larger volumes, the conventional centrifuge is necessary due to the scale limitations of the single-use centrifuge.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 1","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145353551","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

Automation of an integrated micro-scale platform for monoclonal antibody process development by incorporation of a depth filter mimic 单克隆抗体工艺开发集成微尺度平台的自动化，采用深度过滤器模拟。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-16 Epub Date: 2025-10-03 DOI: 10.1002/btpr.70077

Paras Sharma, Petra Sebastian, Lars Robbel, Michael Schmitt, Daniel G. Bracewell

{"title":"Automation of an integrated micro-scale platform for monoclonal antibody process development by incorporation of a depth filter mimic","authors":"Paras Sharma, Petra Sebastian, Lars Robbel, Michael Schmitt, Daniel G. Bracewell","doi":"10.1002/btpr.70077","DOIUrl":"10.1002/btpr.70077","url":null,"abstract":"High throughput process development (HTPD) has been widely adopted for efficient development and optimization of chromatographic operations in monoclonal antibody (mAb) purification. However, the integration of non-chromatographic unit operations, particularly depth filtration following protein A chromatography, which is essential for the removal of process- and product-related impurities prior to the ion exchange chromatography (IEX) operations, remains a challenge due to the absence of commercially available micro-scale depth filtration tools. This limits the integration of this unit operation within the purification sequence, restricting the analysis of process interactions and overall process understanding. In this study, a micro-scale HTPD platform was designed and evaluated to enable integration of a depth filtration mimic, Sartobind® Q anion exchange adsorber, within a mAb purification sequence. This was achieved by translating laboratory-scale protocols to the micro-scale using workflow design tools and executed on an automated liquid handling system. Step yields and impurity clearance were assessed to confirm the equivalence of scale-down. The Sartobind® Q membrane achieved effective removal of host cell DNA (hcDNA), while subsequent IEX operations removed host cell proteins (HCPs) and high molecular weight components (HMWC), meeting target product quality specifications. The platform demonstrated robustness across varying impurity profiles, supporting its applicability for diverse process intermediates. Comparative analysis with laboratory-scale operations confirmed the performance and scalability of the micro-scale system, reducing the total run time by greater than 50%. The integrated HTPD platform offers a resource-efficient, scalable approach for comprehensive mAb purification process development and is suitable for developability assessments during early-stage development.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 1","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12908103/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145211385","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

Dynamic pH profiles drive higher cell-specific and volumetric productivity 动态pH曲线驱动更高的细胞特异性和体积生产力。

IF 2.5 3区生物学

Biotechnology Progress Pub Date : 2026-02-16 Epub Date: 2025-10-20 DOI: 10.1002/btpr.70080

Stephanie R. Klaubert, Dylan G. Chitwood, Danqia Peng, Erin Redman, Ji Young L. Anderson, Nicholas R. Sandoval, Sarah W. Harcum

{"title":"Dynamic pH profiles drive higher cell-specific and volumetric productivity","authors":"Stephanie R. Klaubert, Dylan G. Chitwood, Danqia Peng, Erin Redman, Ji Young L. Anderson, Nicholas R. Sandoval, Sarah W. Harcum","doi":"10.1002/btpr.70080","DOIUrl":"10.1002/btpr.70080","url":null,"abstract":"Mammalian cell cultures in bioreactors rely heavily on critical process parameter control to ensure optimal growth, productivity, and reproducibility to produce recombinant therapeutic proteins. Culture pH has been shown to be a critical parameter that influences growth, productivity, and critical quality attributes. Typically, pH is either controlled to a set-point throughout the culture or uses a single pH shift to achieve higher productivity and more desirable charge variant profiles. The pH is usually maintained by CO2 and base additions. For CO2 controlled cultures, using a set-point can result in an accumulation of CO2, which has detrimental effects on mammalian cell growth and protein production. In this study, a dynamic pH profile was implemented that allowed the pH control in the bioreactor to mimic the natural uncontrolled pH profile observed in shake flask cultures. This dynamic pH profile employs multiple pH shifts during the exponential phase of a single IgG1 producing CHO-K1 cell line. The results show that a dynamic pH profile was able to successfully alleviate CO2 accumulation and increase the cell-specific, as well as overall culture productivity. Impacts of the dynamic pH profile on product quality attributes, including glycosylation and charge variants, were also evaluated, showing mixed impacts on the glycosylation pattern and a positive impact on charge variants. Since the ideal glycosylation pattern is highly dependent on the intended function of the recombinant antibody, impacts on product quality should be evaluated on a “per process” basis.","PeriodicalId":8856,"journal":{"name":"Biotechnology Progress","volume":"42 1","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12908104/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336388","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