Biotechnology and Bioengineering最新文献

{"title":"Model-Enabled Knowledge Transfer Across Cell Lines, Culture Scales and Conditions","authors":"Luxi Yu,&nbsp;Antonio del Rio Chanona,&nbsp;Cleo Kontoravdi","doi":"10.1002/bit.70269","DOIUrl":"10.1002/bit.70269","url":null,"abstract":"<p>Mechanistic models are central to quantitative understanding and optimization of Chinese hamster ovary (CHO) cell culture processes, but their utility is often restricted by parameter sets calibrated for specific cell lines, scales, or operating conditions. In this study, we present the application of the ensemble Kalman filter (EnKF) to bioprocessing, introducing an ensemble-based framework for dual state and parameter estimation that enables mechanistic model adaptation across distinct systems. The EnKF recursively assimilates process measurements to update uncertain kinetic parameters and predict system states. This allows a model calibrated for one system to be transferred to another without reparameterization, using only a single experimental dataset. The evolving parameter ensembles provide a time-resolved sensitivity analysis that identifies which parameters have dominant influence under new process conditions and when their effects become significant. The framework was evaluated across six CHO cell datasets spanning different scales, cell lines, temperatures, and feeding strategies. It accurately reconstructed system dynamics and showed progressive improvement in long-term predictions as data accumulated. By maintaining full mechanistic transparency while flexibly adapting to new data, the EnKF offers a practical route for knowledge transfer across systems, strengthening the role of mechanistic modeling in data-informed bioprocess understanding and control.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2613-2628"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70269","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148263332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Glycoengineered Recombinant Alpha1-Antitrypsin Results in Comparable In Vitro and In Vivo Activities to Human Plasma-Derived Protein","authors":"Frances Rocamora,&nbsp;Chen-Lin Hsieh,&nbsp;Sanne Schoffelen,&nbsp;Johnny Arnsdorf,&nbsp;Eric A. Toth,&nbsp;Abdul S. Yunus,&nbsp;Thomas E. Cleveland IV,&nbsp;Sara Petersen Bjørn,&nbsp;Mina Ying Min Wu,&nbsp;Noel G. McElvaney,&nbsp;Bjørn Gunnar Rude Voldborg,&nbsp;Thomas R. Fuerst,&nbsp;Nathan E. Lewis","doi":"10.1002/bit.70285","DOIUrl":"10.1002/bit.70285","url":null,"abstract":"<p>Alpha-1-antitrypsin (A1AT) is a multifunctional, clinically important, high-value therapeutic glycoprotein that can be used for the treatment of many diseases, such as A1AT deficiency, diabetes, graft-versus-host disease, cystic fibrosis, and various viral infections. Currently, the only U.S. food and drug administration-approved treatment for A1AT disorders is intravenous augmentation therapy with human plasma-derived A1AT (pdA1AT). In addition to its limited supply, this approach poses a risk of infection transmission, since it uses therapeutic A1AT harvested from donors. To address these issues, we sought to generate recombinant human A1AT (rhA1AT) that is comparable to its plasma-derived counterpart using glycoengineered Chinese Hamster Ovary (geCHO-L) cells. By perturbing nine key genes that are part of the CHO glycosylation machinery and expressing the human ST6GAL1 and A1AT genes, we obtained stable, high producing geCHO-L lines that produced rhA1AT having a highly similar glycoprofile to pdA1AT. Additionally, the rhA1AT demonstrated in vitro activity and in vivo half-life comparable to commercial pdA1AT. Thus, we anticipate that this platform will help produce human-like recombinant plasma proteins, thereby providing a more sustainable and reliable source of therapeutics that are cost-effective and better-controlled regarding purity, clinical safety, and quality.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2605-2612"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70285","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148359502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bridging Organ-on-a-Chip and Omics: A Multi-Dimensional Frontier in Biomedical Research","authors":"Zhaoming Cheng,&nbsp;Chuanjun Zhang,&nbsp;Xuwen Li,&nbsp;Yanxue Wei,&nbsp;Zhewei Zhang,&nbsp;Mohan Li,&nbsp;Qi Yu,&nbsp;Yuxin Fang,&nbsp;Di Zhang","doi":"10.1002/bit.70280","DOIUrl":"10.1002/bit.70280","url":null,"abstract":"<div>\u0000 \u0000 <p>Organ-on-a-Chip (OOC) technology offers a powerful platform for replicating human tissue-specific microenvironments, thereby narrowing the translational gap between conventional biomedical models and actual human physiology. Concurrently, omics technologies deliver comprehensive molecular-level insights into biological systems. This review highlights the transformative potential of integrating OOC platforms with high-throughput omics methodologies. We systematically examine the classification, structural configurations, and engineering principles underlying OOC systems, alongside the defining attributes of key omics domains—genomics, transcriptomics, proteomics, and metabolomics. The convergence of dynamic OOC models with advanced omics technologies enables high-resolution, multi-dimensional analyses across numerous biomedical applications, including drug metabolism, disease mechanisms, environmental toxicity assessments, and host-microbiome interactions. This interdisciplinary integration is driving a paradigm shift in precision and translational medicine. However, several challenges remain to be addressed, such as the development of whole-organ mimetics, adaptation of sample collection techniques, and real-time artificial intelligence-based integration of biosensor data with multi-omics datasets. Addressing these hurdles will be vital for unlocking the full potential of this technological synergy in biomedical science.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2474-2490"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148388746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combinatorial Metabolic and Process Engineering for Enhanced GA₄₊₇ Production and Tunable GA₄/GA₇ Ratio in Fusarium fujikuroi.","authors":"Jing-Wen Jia, Zheng-Bin Zhang, Ming-Han Li, Qi-Yun Ma, Lian-Jie Tan, Hao-Yang Li, Zhi-Qing Huang, Yu-Ke Cen, Zhi-Qiang Liu, Yu-Guo Zheng","doi":"10.1002/bit.70380","DOIUrl":"https://doi.org/10.1002/bit.70380","url":null,"abstract":"<p><p>Gibberellins (GAs) are ubiquitous phytohormones that regulate plant growth and are widely used in agriculture. Among these, GA₃ and GA₄₊₇ are the only commercially available products, yet GA₄₊₇ commands a much higher price than GA₃, primarily due to its low titer in the industrial fungus Fusarium fujikuroi. To engineer a high-yielding GA₄₊₇ producer, we first deleted p450-3 to block the conversion of GA₄ and GA₇ to GA₁ and GA₃. This led to the accumulation of GA₄₊₇ at 0.934 g/L, a 37-fold increase over the wild-type strain, albeit with over half reduction in total GA accumulation. Using this Δp450-3 mutant as a platform, we combined metabolic engineering (overexpressing rate-limiting enzymes) with process optimization (pH, medium composition and fermentation duration). These combinatorial interventions synergistically boosted GA₄₊₇ production. Under optimized conditions, the engineered strain achieved a final titer of 6.08 g/L (a 6.51-fold increase over the Δp450-3 parent), comprising 2.13 g/L GA₄ and 3.96 g/L GA₇, representing 5.30- and 7.44-fold increases, respectively. Preliminary optimization in a 10 L fermenter yielded 3.51 g/L of GA₄₊₇. Finally, a solid-state fermentation system was developed on wheat bran, yielding 17.34 g GA₄₊₇ per kg and enabling green, in-house, in-situ gibberellin production. In addition to substantially increasing the GA₄₊₇ titer and total GA accumulation, this study demonstrated that the GA₄/GA₇ ratio can be modulated through both molecular and fermentation strategies.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148962962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering the Vero Cell Lineage: Toward a Programmable Vaccine Manufacturing Platform","authors":"Hanfu Zhang, Zihao Wang, Zhaoqing Yang","doi":"10.1002/bit.70374","DOIUrl":"https://doi.org/10.1002/bit.70374","url":null,"abstract":"Vero cells remain an indispensable continuous substrate for human viral vaccine manufacturing. Despite decades of empirical process optimization, intrinsic genomic instability, including segmental aneuploidy and dynamic chromatin rearrangements, continues to limit the durability of engineered phenotypes under sustained viral burden and bioreactor stress. Here, we review the expanding engineering toolkit for the Vero lineage across a three‐layered functional framework: the membrane interface, cytoplasmic foundry, and nuclear blueprint, evaluating translational prospects at each level. Receptor transplantation and morphological reprogramming have broadened viral entry range and enabled suspension‐adapted culture formats, while metabolic flux management and temporally controlled apoptosis modulation have addressed intracellular production bottlenecks, albeit often with trade‐offs between productivity, biosafety, and long‐term population stability. At the genomic level, targeted perturbations of transcriptional regulators and emerging epigenetic interventions offer more durable gains, yet expression drift, clonal heterogeneity, and karyotypic instability during extended passaging highlight the need for locus‐level precision rather than constitutive trait installation. Looking forward, infection‐responsive dynamic logic circuits and the systematic identification of Vero‐specific genomic safe harbors could shift the paradigm toward a conditionally responsive manufacturing architecture. Collectively, these advances suggest a pathway for transitioning the Vero lineage from a passive, empirically optimized biological substrate into a conditionally responsive, genomically stable, and programmable platform for modern vaccine preparedness.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"51 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148915644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Autonomous Control of Gene Expression Using Endogenous Transcription Signals","authors":"Pakpoom Subsoontorn, Drew Endy, Olivier Borkowski","doi":"10.1002/bit.70377","DOIUrl":"https://doi.org/10.1002/bit.70377","url":null,"abstract":"<jats:label/> Autonomous cell‐based control of heterologous gene expression can simplify batch‐culture bioprocessing by eliminating external monitoring and extrinsic control of culture conditions. Existing approaches use auto‐induction media, synthetic cell‐cell communication systems, or application‐specific biosensors. A simpler, resource‐efficient, and general‐purpose expression control system responsive to common changes during batch culture would be highly valuable. We used native <jats:italic>E. coli</jats:italic> promoters, including P <jats:italic>hdeA</jats:italic> , P <jats:italic>dpS</jats:italic> , P <jats:italic>fumA</jats:italic> , P <jats:italic>rpoA</jats:italic> , P <jats:italic>rpoS</jats:italic> , P <jats:italic>gadA</jats:italic> , and P <jats:italic>yiaG</jats:italic> , and recombinase‐based switches to repurpose endogenous transcription signals for control of heterologous gene expression. Specifically, natural changes, covering 1–2 orders of magnitude across growth phases, in transcription from endogenous promoters result in recombinase expression at the exponential‐to‐stationary phase transition. So‐expressed recombinases invert a constitutive promoter regulating expression of arbitrary heterologous genes amplifying the endogenous transcriptional input signal by more than 10‐fold. We realized reversible and single‐use switching with reduced static and dynamic cell‐to‐cell variation and overall expression amplification. We used “off‐the‐shelf” genetic parts and abstraction‐based composition frameworks to realize reliable forward engineering of our synthetic genetic systems. We engineered autonomous control systems for regulating heterologous gene expression. Our system uses generic endogenous promoters to sense and control heterologous expression during growth‐phase transitions. Our system does not require specialized auto‐induction media, production or activation of quorum sensing, or the development of application‐specific biosensors. Cells programmed to control themselves could simplify existing bioprocess operations and enable the development of more powerful synthetic genetic systems. Modern biotechnology uses engineered microbes to manufacture molecules that are incorporated into medicines, flavors, fuels, and other materials. Making molecules inside cells requires enzymes, whose expression levels and timing need to be optimized to maximize final product yields. Many engineered microbes use enzymes that should only be expressed during the later stages of cell growth to avoid overconsumption of resources or accumulation of toxic intermediates. Here, we develop simple genetic devices that enable engineered cells to control themselves, automatically switching on or off enzyme expression during batch cell culture. Unlike existing approaches, our devices do not require specialized growth media or wasteful synthesis of cell‐cell signaling molecule.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"6 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148915645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cross-Kingdom Siderophores: Biosynthesis, Ecology, and Biotechnological Applications.","authors":"Debapriya Maitra, Aadishri Pai, Shah Qamar, Soumyadeep Chakraborty, Shareen Singh, Harjot Singh Gill, Mithul Rajeev, Soumya Pandit, Moupriya Nag, Dibyajit Lahiri","doi":"10.1002/bit.70367","DOIUrl":"https://doi.org/10.1002/bit.70367","url":null,"abstract":"<p><p>Microbial siderophores are high-affinity iron-binding compounds which are produced by bacteria, fungi, and actinomycetes to obtain iron and survive and interact with different species in an iron-deficient environment. While the conventional research on siderophore systems deals mainly with the study within the same taxa, modern researchers have increased their inclination toward cross-kingdom integration of siderophore behavior and their impact on host-associated environments. This can be largely attributed to differences in biosynthetic gene clusters, receptor systems, and regulatory networks, which produce distinct genotype-to-phenotype results determining microbial cooperation and competition. Current advancements in genomic research, together with omics studies like transcriptomics, proteomics, and metabolomics, have created newer insights into how siderophores function. However, the present literature evidences multiple major gaps in multi-omics data because the link between genomes and metabolomes remains weak due to inconsistent regulatory data sets and failure in identifying producer-consumer relationships in polymicrobial systems. Additionally, major constraints like molecular instability, delivery system limitations, host toxicity, limitations in upscaling, and regulatory issues delimit the use of siderophores in medical treatment, agricultural practices, and environmental biotechnology. This review aims to bridge the existing knowledge about siderophore biochemistry, biosynthesis, ecological functions, and genetic regulation across kingdoms while integrating multi-omics outlook with translational considerations. Thus, by connecting molecular mechanisms with evolutionary cross-talk, this study aims to provide a system-level framework in the world of siderophore-mediated iron uptake and therefore shapes future directions in emerging fields of microbial engineering, precision therapies, and sustainable biotechnology.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148900457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Architectural Design of Phage-Displayed Peptide Libraries for Peptide Discovery.","authors":"Belle Li Ling, Huan Peng, Theam Soon Lim","doi":"10.1002/bit.70370","DOIUrl":"https://doi.org/10.1002/bit.70370","url":null,"abstract":"<p><p>Peptide phage display is a cornerstone technology in molecular engineering, providing a robust platform for the rapid discovery of high-affinity ligands. The rapid rise of peptides, especially in therapeutics, over recent years has shed light on the importance of phage display as a tool for peptide discovery. The evolution of peptide scaffolds from flexible linear sequences to structurally constrained architectures has expanded the scope of application of peptides in general. The review examines strategies used to generate monocyclic, bicyclic, and multicyclic peptides. These constrained formats mimic the complex loops of natural proteins, offering enhanced metabolic stability and superior binding affinity for challenging targets like protein-protein interaction interfaces. Furthermore, the review surveys the diverse applications ranging from targeted therapeutics and molecular diagnostics to the engineering of material-specific peptides for nanotechnology. Finally, we discuss current technical hurdles and the transformative potential of integrating next-generation sequencing and computational modeling to redefine the future of peptide discovery.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148900422","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Green Chemoenzymatic Route to 28-epi-Homobrassinolide Enabled by OsO₄-Free Strategy and Engineered Baeyer-Villiger Monooxygenase With MOF Immobilization.","authors":"Chuanyu Fu, Ruoyu Jiao, Yunyijie Zhou, Yamei Lin, Xing Zhang","doi":"10.1002/bit.70373","DOIUrl":"https://doi.org/10.1002/bit.70373","url":null,"abstract":"<p><p>28-epi-homobrassinolide (28-epi-HBL), a bioactive analog of the steroidal phytohormone brassinolide, exhibits broad biological activities and considerable application potential. However, conventional chemical routes remain limited by tetrahydroxylation step that relies on highly toxic osmium tetroxide in combination with costly chiral ligands, whereas the Baeyer-Villiger oxidation step requires strong oxidants and often exhibits limited regioselective control. In this work, optimization of tetrahydroxylation was achieved using the Shing-Plietker reaction, thereby avoiding the use of osmium tetroxide. Moreover, an engineered Baeyer-Villiger monooxygenase (CpnB_BL) was heterologously expressed and used for regioselective B-ring lactonization under mild conditions. A ZIF-8-based immobilization strategy was established for this engineered enzyme, further improving its stability and solvent tolerance, and increasing the isolated yield of the Baeyer-Villiger step to 83%. These optimized strategies were subsequently applied to the total synthesis of 28-epi-HBL, achieving an overall yield of 44.9%. This represents a marked improvement over the highest previously reported yield in the literature, which stands at approximately 20%. Collectively, this study lays the groundwork for the development of more precise and sustainable synthetic routes to 28-epi-HBL.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148900495","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microfiltration (MF) and Ultrafiltration (UF) for Sewage and Surface Water Concentration for SARS-CoV-2 and Other Enveloped Viruses' Detection in Wastewater-Based Epidemiology (WBE).","authors":"Rodrigo Almeria Ragio, Rodrigo de Freitas Bueno, Roseli Frederigi Benassi, Eduardo Lucas Subtil","doi":"10.1002/bit.70368","DOIUrl":"https://doi.org/10.1002/bit.70368","url":null,"abstract":"<p><p>This study objective was to evaluate hollow fiber membranes for concentration of enveloped enteric viruses in sewage and surface water samples. Microfiltration (MF, 0.2 µm nominal pore size) and ultrafiltration (UF, 100 kDa molecular weight cut-off) hollow fiber membranes were compared for the concentration of the bovine respiratory syncytial virus, and the influence of initial sample volumes (1, 1.5, and 2 L), permeate recovery rates (60%, 70% and 80% of the initial volume collected as permeate, equivalent to volumetric concentration factors of 2.5, 3.3 and 5), and backwashing was assessed. Polyethylene glycol precipitation followed by centrifugation (PEG-C) was evaluated both as a stand-alone method and as a secondary step applied to the membrane concentrate. MF and UF membranes had a similar capacity to retain pollutants and to concentrate organic matter in most parameters and conditions (p > 0.05). In sewage, increasing the initial volume from 1 to 2 L raised recovery from 20.7 ± 0.1% to 30.0 ± 0.1% with MF and from 20.8 ± 0.1% to 25.3 ± 0.1% with UF (p < 0.05), whereas changes in the permeate recovery rate produced no significant differences (p > 0.05). In surface water recovery did not exceed 5.5%, and every increment in the permeate recovery rate produced a significant increase (p < 0.05). Backwashing had a stronger effect on RNA recovery than pore size or the remaining operational parameters, reaching 29.3 ± 9.3% with MF and 33.8 ± 13.7% with UF in sewage and nearly doubling recovery in surface water. In sewage, PEG-C alone (14.1%) was statistically similar to membrane filtration alone (12.1% to 12.3%, p > 0.05), while combining both techniques raised recovery to 23.0% to 25.5%. In surface water, membranes alone recovered 0.8% against 2.4% for PEG-C (p < 0.05), and the combination of UF with PEG-C reached 5.7%, making it a valid alternative, albeit more time consuming and requiring additional equipment. Recoveries are expressed as the ratio between the genome copy concentration in the concentrate and in the spiked sample; on a total genome copy basis they correspond to 4% to 9% in sewage and to about 1% in surface water.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148886446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}