Biotechnology and Bioengineering最新文献

{"title":"Critical Review on Microbial Inulinase Production: Emerging Strategies, AI-Driven Optimization, and Applications","authors":"Sagar S. Dhande,&nbsp;Nikita A. Mankoskar,&nbsp;Harris P. Panakkal,&nbsp;Indarchand R. Gupta,&nbsp;Rahul P. Bhagat","doi":"10.1002/bit.70306","DOIUrl":"10.1002/bit.70306","url":null,"abstract":"<div>\u0000 \u0000 <p>Microbial inulinases are increasingly recognized as valuable biocatalysts for the sustainable production of high-value products, including fructooligosaccharides, fructose, bioethanol, and organic acids in industries, such as food, pharmaceuticals, and bioenergy. In the last few decades, microbial inulinase research has advanced significantly, from strain selection and fermentation optimization to advanced enzyme engineering and immobilization, improving yields, stability, and reusability. There are still some final bottlenecks, such as low yields, poor thermostability, and high purification costs. This review examines strategies to innovate and overcome these bottlenecks, including novel immobilization strategies that utilize nanomaterials, system-scale bioprocess optimization using artificial intelligence (AI), and bioprospecting extremophiles using metagenomics. The present review discusses how statistical and computational modeling (RSM, ANN, and AI) significantly increases yield and process efficiency, with comments on their relevance to contemporary biorefinery applications. The advanced immobilization approaches significantly enhance operational stability and reusability, allowing for continuous processing. This review situates the development of inulinase as not just an enzymological effort but a multidisciplinary effort involving process engineering and sustainability science. Overall, emphasize is given toward the thought that advancements leaning toward the future will require a synthesis of AI-designed enzyme systems; economical immobilization supports; and incorporation of circular bioeconomy principles through the valorization of agro-wastes. These barriers to knowledge transfer must be resolved if we are to unlock the full bioeconomic potential of microbial inulinase systems.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"2856-2883"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148428811","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":"Current Applications of Staphylococcal Protein A in Biotechnology and Biomedicine: A Comprehensive Review","authors":"Shirin Damough,&nbsp;Shadi Damough,&nbsp;Ladan Mafakher,&nbsp;Masoumeh Heshmati,&nbsp;Ahmad Adeli,&nbsp;Yeganeh Talebkhan","doi":"10.1002/bit.70307","DOIUrl":"10.1002/bit.70307","url":null,"abstract":"<div>\u0000 \u0000 <p>Protein A (SpA), a cell wall-associated protein of <i>Staphylococcus aureus</i>, is an immunoglobulin (Ig) binding molecule with high affinity to the Fc moiety of IgG. It can help <i>S. aureus</i> for effective shielding from recognition and opsonization by immune cells, thereby promoting bacterial immune evasion. Since its recognition in the early 1980s, diverse applications have been developed across biotechnology, medicine, and biochemical research, using both native and recombinant SpA variants produced in various host cells. This review provides a comprehensive overview of the major applications of SpA, with an emphasis on techniques that progressed to commercially available tools/platforms. These applications are based on its Ig-binding capability, including purification of monoclonal antibodies in biopharmaceutical manufacturing, selective isolation of antibodies in diagnostic workflows, integration into serological assays or biosensor systems, and use as a fusion partner/tag for capturing recombinant proteins.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"2884-2905"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148521910","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 Stable Lentiviral Vector Producer Cells Improves LVV Production","authors":"Jona Röscheise,&nbsp;Lena-Marie Eberle,&nbsp;Holger Laux,&nbsp;Kerstin Otte","doi":"10.1002/bit.70318","DOIUrl":"10.1002/bit.70318","url":null,"abstract":"<p>Stable lentiviral vector producer cell lines represent a promising platform for scalable and cost-efficient vector manufacturing, yet their productivity is often limited by intrinsic host-cell constraints. In this study, we aimed to identify cellular factors restricting LVV production in stable doxycycline-inducible GPRTG producer cell lines and to evaluate whether targeted host-cell engineering can improve vector yield. Comparative transcriptomic analysis of high- and low-producing clones revealed distinct differences in cellular pathways related to transcription, translation, energy metabolism, lipid homeostasis, stress response, and chromatin regulation. Based on differential gene expression, candidate genes were modified by CRISPR/Cas9-mediated knockout or overexpression and functionally analyzed in a low-producing clone. Single-gene KO screening identified H1-2, ADAMTS1, INSIG1, GADD45B, and HSPA1B to increase cell-specific LVV productivity up to 2.6-fold, with H1-2 showing the strongest effect. In addition, combinatorial disruption further improved productivity. In contrast, overexpression of selected candidates did not enhance LVV production. Overall, our results demonstrate that transcriptomics-guided host-cell engineering is an effective strategy to identify and relieve intrinsic bottlenecks in stable LVV producer cell lines.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3152-3164"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70318","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148521914","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":"Synergistic Co-Fermentation of Ulva and Waste Activated Sludge for Sustainable Resource Recovery: Mechanistic Insights From Dissolved Organic Matter Transformation and Multi-Omics Analysis.","authors":"Lifeng Wang, Yibo Zhang, Xinyan Zou, Yazhi Nong, Chenyuan Dang, Jie Fu","doi":"10.1002/bit.70410","DOIUrl":"https://doi.org/10.1002/bit.70410","url":null,"abstract":"<p><p>Anaerobic fermentation for volatile fatty acids (VFAs) production represents the sustainable and resource-recovery strategy for waste activated sludge (WAS) treatment, yet its efficiency is often hampered by low substrate C/N ratio and limited hydrolysis. Ulva, a macroalga responsible for extensive \"green tides\" that burden coastal ecosystems and economies, emerges as an ideal co-substrate for WAS anaerobic fermentation due to its high C/N ratio and biodegradability. This study established a novel Ulva-WAS co-fermentation system to boost VFAs production and explained the mechanism of acidogenic enhancement and methanogenesis suppression through integrated analyses of VFAs generation, dissolved organic matter (DOM) molecular transformation, and microbial metabolic activity. Co-fermentation enhanced VFAs production (1.67-2.9-fold increase of maximum VFAs accumulation) while stimulating microbial activity and accelerating hydrolysis of macromolecular components into polysaccharides and amino acids. Within the carbohydrate-supplemented environment, DOM transformation was dominated by microbial acidogenic processes, including glycolysis and amino acid metabolism. Microbial community succession and metabolic profiling confirmed that the high-C/N, carbohydrate-rich fermentation environment drove deterministic assembly of communities toward the function of acidogenesis, which in turn further promoted VFAs production. Concurrently, reactive oxygen species (ROS) stress induced by VFAs accumulation selectively inhibited methanogenic activity. This work provides an innovative strategy for WAS treatment and offers profound insights into the interplay between DOM dynamics and microbial metabolism in co-fermentation systems.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149165524","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":"From Sequence to Structure: AlphaFold and the AI Revolution in Aquaculture Drug Discovery and Vaccine Design.","authors":"Enes Üstüner, Mustafa Öz","doi":"10.1002/bit.70398","DOIUrl":"https://doi.org/10.1002/bit.70398","url":null,"abstract":"<p><p>Global aquaculture faces an escalating disease and antimicrobial resistance (AMR) crisis, exacerbated by climate-driven physiochemical shifts and intensive farming practices. The integration of artificial intelligence (AI) and structural biology, spearheaded by AlphaFold, represents a paradigm shift in aquatic pharmacology, transitioning the field from empirical trial-and-error to rational in silico design. This review critically evaluates the application of AI-driven tools in aquaculture drug discovery and vaccine development. We examine the evolution of AlphaFold in predicting complex host-pathogen interactions and facilitating de novo drug design. Concurrently, we highlight critical limitations stemming from the reliance on multiple sequence alignment (MSA) depth rather than mere PDB taxonomic composition, the unreliability of direct molecular docking due to apo conformations, and the dynamic nature of psychrophilic adaptations in aquatic environments. Furthermore, we assess next-generation structural vaccinology, including reverse vaccinology and pre-fusion antigen stabilization, alongside the integration of nanomaterial-based delivery systems to overcome aquatic pharmacokinetic barriers. The synergy of AI with CRISPR/Cas technologies and multimodal large language models (MM-LLMs) for disease management is also explored. Despite unprecedented computational capabilities, significant bottlenecks remain, including a severe lack of species-specific omics data, the indispensability of in vivo wet-lab validation, and lagging regulatory frameworks. Ultimately, realizing the potential of precision aquatic medicine requires developing aquaculture-specific databases, addressing the ecological risks of open-water deployment, and democratizing these technologies for small-scale farmers to ensure sustainable, equitable, and resilient global aquaculture.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149164918","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":"Efficient Cell Removal and Impurity Reduction for Poxvirus Purification in a Single Step Alluvial Filtration.","authors":"Linus G Weber, Caroline Stephan, Georgios Makris, Leander John, Ingo Jordan, Michael W Wolff","doi":"10.1002/bit.70413","DOIUrl":"https://doi.org/10.1002/bit.70413","url":null,"abstract":"<p><p>Modified Vaccinia virus Ankara (MVA) is a potent vector for the development of vaccines and novel cancer treatment strategies. Since it is a non-replicating virus in human recipients, high infective titers are required for therapeutic efficacy. Harvesting and clarification of cell culture-derived MVA are critical for the overall process yield and remains challenging, as full MVA release depends on cell lysis, resulting in a difficult-to-filter cell lysate. In this study, we present the development of a high-yield clarification strategy using diatomaceous earth (DE) as a natural filter aid, followed by an optimized endonuclease treatment. An ionic strength containing a 650 mM salt mixture of NaCl, NaBr, and KCl allowed impurity depletion by DE binding while maintaining a complete virus recovery. The optimal alluvial filtration conditions were achieved with a PURAFIX® CH 09 depth filter with a 10-30 µm retention rate. The optimal conditions were identified as a combination of a Celpure® C1000 precoat and a Celpure® C300 additive added to the feed. This robust process achieved 94% total virus recovery while reducing turbidity to 23% and residual DNA and protein impurities to 76% and 72%, respectively, in a single-step filtration. Subsequent nuclease digestion benefits from a reduced particle and impurity burden, resulting in further 93% DNA depletion under optimal conditions of 11 h of incubation at 21°C with 51 U/mL of the salt active DENARASE® High Salt endonuclease. The study demonstrated that sterilized filter aids enable a single-step filtration to recover high MVA titers with a reduced impurity burden, paving the way for improved downstream process efficiency.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149164150","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":"Advanced In Vitro Platforms for Experimental Liver Research: From Organoids and Liver-on-Chip to Omics and Artificial Intelligence.","authors":"Seyedeh Kiana Teymorian, Pedram Asadi Sarabi, Abbas Piryaei, Massoud Vosough","doi":"10.1002/bit.70411","DOIUrl":"https://doi.org/10.1002/bit.70411","url":null,"abstract":"<p><p>Hepatocytes perform essential physiological functions in the human body. Gaining more profound insight into the liver structure, diseases, and physiology by using in vitro models can help to propose more relevant clinical applications. Conventional in vitro liver models, such as two-dimensional (2D) cell cultures, fail to adequately replicate the hepatic microenvironment, even when improved by adding additional cell types and extracellular matrix (ECM). These models do not address all necessary key components to mimic in vivo conditions, particularly multicellularity, specific ECM, and proper biophysical and biochemical parameters. Organoids, 3D-bioprinted constructs, and liver-on-chip platforms establish physiologically relevant models by improving cell-cell and cell-matrix interactions and multicellular architecture. For a comprehensive understanding of the liver's interconnections with related tissues and organs, a multi-organ-on-chip is a useful platform for more systemic studies. Also, other interdisciplinary approaches, particularly omics and artificial intelligence (AI), can enhance the accuracy of experimental and pre-clinical data achieved by such advanced in vitro liver platforms. This review highlights developing platforms for integrated approaches in experimental liver studies to enable researchers to have a holistic view of the liver in physiologic and pathologic conditions.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149164386","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":"Development of Immobilized Restriction Enzyme Reactors on a Monolith Backbone for Streamlined Plasmid DNA Linearization.","authors":"Maša Nabergoj, Špela Peršič Debenjak, Polona Skrt, Tina Vodopivec Seravalli, Polona Megušar, Manuela Bartolini, Rok Sekirnik, Aleš Štrancar, Sebastijan Peljhan","doi":"10.1002/bit.70406","DOIUrl":"https://doi.org/10.1002/bit.70406","url":null,"abstract":"<p><p>High-quality, properly linearized plasmid DNA (pDNA) is a decisive upstream determinant of in vitro transcription (IVT) performance, impacting mRNA yield, integrity, and batch-to-batch consistency. We developed immobilized restriction enzyme reactors (IMERs) on monolithic supports and benchmarked EcoRI, NdeI, and HindIII to evaluate suitability for IVT-grade template preparation. Across all three enzymes, IMERs quantitatively converted supercoiled and open-circular pDNA to the linear form and delivered a clean product, as verified by baseline-resolved chromatography and agarose gels. To further verify the functionality of the IMER platform, BspQI was immobilized and used to linearize a second plasmid construct. IVT initiated from all four IMER-processed templates produced mRNA with kinetics and titers indistinguishable from conventional in-solution digestion, underscoring that template quality was preserved. Long-term stability studies showed enzyme-dependent stability with HindIII-based IMER retaining ~20% activity for more than 4 months. While the normalized productivity of the IMER matched that of in-solution reactions, the possibility of reusing IMER across multiple linearization cycles significantly increased cumulative yield. Further, linearization by IMERs eliminated the need for post-reaction enzyme removal and harsh inactivation, thereby reducing degradation risk. Collectively, these results establish restriction enzyme-based IMERs as a robust, scalable, automation-ready tool for producing high-purity, IVT-ready linear pDNA, securing the template quality required by the current mRNA manufacturing process.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149165168","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":"Curation of Gene–Protein–Reaction Associations Enhances the i Pichia Genome‐Scale Metabolic Model of Komagataella phaffii","authors":"Pınar Kocabaş","doi":"10.1002/bit.70402","DOIUrl":"https://doi.org/10.1002/bit.70402","url":null,"abstract":"<jats:italic>Komagataella phaffii</jats:italic> is a commercially valuable yeast widely used in biotechnology due to its Crabtree‐negative metabolism, which enables it to sustain respiratory growth and efficiently utilize carbon sources even in the presence of excess glucose. To better understand and predict its metabolic behavior, genome‐scale metabolic models are commonly employed. Within these models, Gene–Protein–Reaction (GPR) associations play a crucial role in the predictive performance of models. The previously developed <jats:italic>i</jats:italic> Pichia genome‐scale metabolic model for <jats:italic>K. phaffii</jats:italic> was further refined in this study through extensive manual curation of GPR associations, resulting in an updated model termed <jats:italic>i</jats:italic> Pichia2. GPR associations were manually curated for 862 reactions based on the enzymatic functions and subcellular localizations of the associated proteins, using information obtained from the KEGG, NCBI, and UniProtKB databases. Gene essentiality analyses were subsequently performed using <jats:italic>i</jats:italic> Pichia2, and the predicted results were compared with experimental data reported in the literature. This refinement of GPR associations improved the overall predictive performance of the model, with prediction accuracy increasing from 0.71 in <jats:italic>i</jats:italic> Pichia to 0.727 in <jats:italic>i</jats:italic> Pichia2. Similarly, the Matthews correlation coefficient (MCC) increased substantially from 0.1075 to 0.24, indicating improved agreement between predicted and experimentally observed gene essentiality phenotypes. Compared with <jats:italic>i</jats:italic> Pichia, <jats:italic>i</jats:italic> Pichia2 predicted fewer true positives (313 vs. 391), false positives (69 vs. 108), and false negatives (67 vs. 71), while the number of true negatives increased from 36 to 49. The higher accuracy and MCC obtained with <jats:italic>i</jats:italic> Pichia2, despite its lower overall MEMOTE score (%34) compared with the original <jats:italic>i</jats:italic> Pichia model (%52), highlights that model quality and predictive performance represent distinct aspects of genome‐scale metabolic model evaluation. Overall, these results demonstrate that manual refinement of GPR associations can improve the predictive performance of GEMs without modifying their underlying reaction network. The improved GPR associations in <jats:italic>i</jats:italic> Pichia2 are therefore expected to support more reliable identification of gene knockout targets and facilitate metabolic engineering and rational strain design in <jats:italic>K. phaffii</jats:italic> .","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"11 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149123865","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":"Cryo‐EM Analysis of Adenovirus Vector Reveals Purification Conditions Leading to Loss of Viral Fiber Proteins and Reduced Infectivity","authors":"Betsy Widdicombe,Joseph D. Barritt,Nichakorn Pipatpadungsin,Braulio Carrillo Sanchez,Dale Stibbs,Lisa Cooper,Sarah L. Rouse,Daniel G. Bracewell","doi":"10.1002/bit.70407","DOIUrl":"https://doi.org/10.1002/bit.70407","url":null,"abstract":"ABSTRACT Vaccines and gene therapies require efficient delivery of genetic material to host cells. Viral vectors leverage viruses' ability to enter host cells to achieve this. Following the SARS‐CoV‐2 vaccine and novel gene therapies that utilize adenovirus vectors, there has been an increase in the demand for large‐scale production of vectors. A benefit of adenovirus vectors is that they can act as a “plug and play” tool for any future emerging diseases for the development of vaccines. A key obstacle was the heterogeneity that is found in the final product. This can reduce the functionality of the vector and affect immune responses. A possible origin of heterogeneity is the downstream processing, specifically the bind and elute steps of anion exchange chromatography (AEX), which is widely used at scale. To better understand the heterogeneity that AEX could elicit on the adenovirus capsid, cryogenic‐electron tomography was used to assess the capsid integrity after a variety of chromatography conditions in comparison to a cesium chloride (CsCl) gradient ultracentrifugation purified control. To investigate the effect of absorption time during chromatography, AEX conditions with a 5, 10, and 20 min on‐resin absorption periods were evaluated. Following AEX binding at pH 6.5, samples showed a reduction from 17% in the CsCl purified control to 10% infectious particles. When examined using electron microscopy, the virions exhibited aggregation and a decrease from 70% to 54% in fiber occupancy. After a 20 min AEX hold at pH 6.5 there was almost complete loss of infectivity and fiber occupancy decreased from ~70% to below 30%. Samples run at pH 8 did not show the same loss in infectivity when held bound for 20 min. This suggests that purification conditions can cause removal of the fiber protein, which was likely causing the decrease in infectivity, due to the fiber's role in cell entry. We hypothesize that fiber removal was caused by a combination of hold time and weakened binding between the fiber and penton base due to the low pH conditions that are often used during AEX. This work demonstrates a link between fiber loss and reduced adenovirus vector functionality and how purification conditions can impact vector functionality. Through studying the relationship between structure and function, we can hopefully minimize loss of infectivity during purification by optimizing processing conditions, reduce manufacturing cost and improve the quality of adenovirus vaccines and gene therapies.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"32 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149090288","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}