Biotechnology and Bioengineering最新文献

{"title":"Pectinase-Based Bioprocesses for Circular Bioeconomy and Sustainable Industrial Development","authors":"Nisha Sharma,&nbsp;Dimple Tanwar,&nbsp;Usha Grewal,&nbsp;Murugesan Mohana Keerthi,&nbsp;Yogesh K. Ahlawat,&nbsp;Nitin Pandey,&nbsp;Chirag G. Makvana","doi":"10.1002/bit.70312","DOIUrl":"10.1002/bit.70312","url":null,"abstract":"<p>Pectinases play a vital role in the degradation of pectic part of the plant cell wall and are considered in the group of hydrolytic enzymes. In the present scenario, demand of economically feasible and environment friendly techniques and approaches has significantly led the research on the microbial production, standardization of process parameters and improvements of pectinases. The development of pectinase-based bioprocesses has been found to promote sustainable industrial practices by reducing the use of chemicals, energy, and waste, thereby supporting eco-friendly production systems. The unique aspect of this review is that it explains traditional methods of pectinase production alongside novel methods, which include solid-state and submerged fermentation, recombinant DNA techniques, heterologous gene expression, protein engineering, CRISPR/Cas genome editing technologies, enzyme immobilization, and nanobiotechnological methods, which improve the production, stability, and performance of pectinases. It further demonstrates the increasing importance of the use of pectinases in sustainable manufacturing through decreasing the use of chemicals and energy, as well as reducing waste production from industry. It illustrates that innovations in microbial strain engineering, process optimization, and utilization of low-cost substrates have significantly increased the economic viability and efficiency of producing pectinases. Yet, some issues associated with commercialization, standardization of production processes, recovery, and stability of enzymes still exist. Overall, this review paper gives an overview of the recent advancements and limitations of next-generation pectinases.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"2906-2936"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70312","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148617980","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":"High Cell Density Cultivation of HEK293 Cells Using a 2L Membrane-Stirred Bioreactor","authors":"Lennart Jacobtorweihe,&nbsp;Yasemin van Heuvel,&nbsp;Kathrina Scheiermann,&nbsp;Patrick Bongartz,&nbsp;Yvonne Genzel,&nbsp;Udo Reichl","doi":"10.1002/bit.70317","DOIUrl":"10.1002/bit.70317","url":null,"abstract":"<p>The use of perfusion systems for the continuous cultivation of animal cells at high cell density (HCD) for the production of recombinant proteins, viruses, and viral vectors has many advantages. Besides an enhancement of volumetric productivity and high cell viability, the product quality can be improved through a steady-state environment. However, the establishment of such a process intensification method using stirred tank bioreactors (STRs) for cultivations at very high cell concentrations (&gt; 50 × 10⁶ cells mL⁻¹) can be challenging. In particular, limitations in the oxygen supply, CO₂ stripping, high viscosity, and excessive foam formation can impede cell viability and product yields. In this study, we investigated a novel membrane-stirrer (MemStir) based bioreactor that employs multiple hollow-fiber membrane sheets arranged in a device that facilitates efficient gas transfer through bubble-free diffusion for HCD cultivations. We found that the oxygen transfer rates at different tip speeds and gas flow rates resulted in volumetric mass transfer coefficient (kLa) values of 10–30 h⁻¹ enabling sufficient oxygen supply to support concentrations exceeding 50 × 10⁶ cells mL⁻¹. The functionality of the membrane-stirrer was benchmarked in batch mode against shake flasks and a conventional STR equipped with a pitched-blade impeller and an <span>l</span>-drilled hole sparger. Suspension growth of a human embryonic kidney 293 LTV (HEK-LTV) cell in the MemStir system was comparable to the cell growth in a STR. Using pure oxygen for aeration, a maximum viable cell concentration of 7.6 × 10⁶ cells mL⁻¹ was achieved. As a starting point for further optimization, we investigated the impact of the MemStir system on viable cell concentration, cell viability, dissolved oxygen partial pressure, pH control and foam formation in HCD cultivations in perfusion mode. Using the MemStir system equipped with an alternating tangential flow system, HEK-LTV cells reached concentrations of up to 92 × 10⁶ cells mL⁻¹ with a specific growth rate of 0.022 h⁻¹ in perfusion mode. Moreover, controlling the pH in the range 7.0 to 7.3 was easily achieved, and no foam formation was observed. Overall, these results suggest that the MemStir system is a viable option for HCD cultivation because it can overcome limitations of both aeration and foam formation.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3165-3174"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70317","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148589518","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":"Secretion Engineering Enables Production of a Structurally Stable and Long-Continuous Type III Collagen Fragment in Pichia pastoris","authors":"Lin Zhu,&nbsp;Qingping Liang,&nbsp;Guijun Zhang,&nbsp;Zhaoyu Wang,&nbsp;Zhaopeng Shen,&nbsp;Xiaodan Fu,&nbsp;Changliang Zhu,&nbsp;Haijin Mou","doi":"10.1002/bit.70311","DOIUrl":"10.1002/bit.70311","url":null,"abstract":"<div>\u0000 \u0000 <p>Recombinant humanized type III collagen has attracted increasing interest for biomedical and tissue engineering applications due to its roles in extracellular matrix remodeling and tissue repair. However, the production of long-continuous, structurally stable type III collagen fragment in <i>Pichia pastoris</i> is limited by host-derived proteolysis of the recombinant α1 chain, resulting in fragmentation and loss of structural integrity. In this study, a long-continuous type III collagen fragment was produced in <i>P. pastoris</i> by proteolytic selection and identified as a continuous 585-amino acid sequence (N611–P1195) by LC–MS/MS. Secretion efficiency was improved by signal peptide optimization, enabling the development of high-producing strains (6.22 g/L). rColIII was purified to &gt; 90% purity with low endotoxin levels (&lt; 10 EU/mg) using multimodal chromatography and exhibited enhanced thermal stability. Although CD analysis indicated the absence of a canonical triple-helix conformation, FITR spectroscopy confirmed preservation of key peptide backbone structures. Biological assays demonstrated favorable cytocompatibility, with rColIII promoting HSF cell viability, adhesion, and migration. This study provides a scalable strategy for producing long, continuous type III collagen fragment with improved stability in <i>P. pastoris</i>, and supports their potential applications in cosmetic, tissue engineering, regenerative medicine, and other biomedical fields.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3106-3117"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148729215","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":"Enhancing the Secretion Systems: Genetic Engineering of Super Bioagents for Effective Plant Disease Control","authors":"Michael Dare Asemoloye","doi":"10.1002/bit.70303","DOIUrl":"10.1002/bit.70303","url":null,"abstract":"<p>The escalating threat of plant diseases to global agriculture and food security necessitates innovative and sustainable control strategies. Conventional biological control agents (BCAs), while environmentally friendly, often suffer environmental challenges and secretion of limited/poor antimicrobial compounds. Advances in CRISPR/Cas genome editing, protease engineering, and synthetic biology have enabled precise modifications that improve pathogen targeting and secretion efficiency. Interest should now be shifted on development of “Super Bioagents (SBs)” with enhanced secretion systems (SSs) for plant disease suppression against changing environmental factors. This will create sustainable ecofriendly alternative to chemical pesticides. This review explores a detailed overview of molecular mechanisms of microbial SSs and the potentials of SBs as a frontier in plant disease management. While there are still challenges in mass deployment of BCAs in sustainable agriculture, this review is guided by the hypothesis that rational, quantitative engineering of microbial SSs can transform conventional BCAs into integrated SBs. It synthesizes current advances within a systems‑level bioengineering framework linking secretion efficiency, regulation, and field performance. It further explores possible integration of SBs in plant–microbiome interactions to further enhance their adaptability and effectiveness. Finally, the review dives into recent breakthroughs, current challenges, and future directions for SBs development and application as next-generation plant disease control agents.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"2821-2855"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70303","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148428791","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":"Biotechnology and Bioengineering: Volume 123, Number 11, November 2026","authors":"","doi":"10.1002/bit.70418","DOIUrl":"https://doi.org/10.1002/bit.70418","url":null,"abstract":"","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"2817-2820"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70418","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149177382","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":"Developing Anti-EGFR/Anti-HER2 Bifunctional Antibody for Solid Tumors by Protein Engineering","authors":"Senem Sen,&nbsp;Aslı Semerci,&nbsp;Recep Erdem Ahan,&nbsp;Cemile Elif Ozcelik,&nbsp;Elif Duman,&nbsp;Behide Saltepe,&nbsp;Ebru Sahin Kehribar,&nbsp;Ebru Aras,&nbsp;Eray Ulas Bozkurt,&nbsp;Hilal Yazıcı,&nbsp;Müge Serhatlı,&nbsp;Urartu Ozgur Safak Seker","doi":"10.1002/bit.70304","DOIUrl":"10.1002/bit.70304","url":null,"abstract":"<div>\u0000 \u0000 <p>Overexpression of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) is common in solid tumors like breast, colorectal, and head and neck cancers, driving oncogenic signaling, therapy resistance, and poor prognosis. Monotherapies often fall short in fully suppressing tumor progression, particularly in cases with EGFR/HER2 co-amplification. Dual-targeting strategies offer enhanced efficacy by mitigating resistance and amplifying antitumor responses. Our study focused on developing and characterizing a bispecific anti-EGFR/HER2 antibody designed to simultaneously block ligand binding and receptor activation while harnessing Fc-mediated immune effector functions. Utilizing knob-into-hole and CrossMab technologies, we engineered a bispecific antibody capable of binding the extracellular domains of both EGFR and HER2. <i>In vitro</i> analyses confirmed its dual-binding capacity. Comparative studies revealed that this bispecific construct outperformed its monospecific counterparts in suppressing proliferation and, notably, in increasing the expression of apoptotic markers in EGFR/HER2-expressing tumor cells. Together, these findings highlight the synergistic therapeutic potential of bispecific antibodies as a promising modality for overcoming the limitations of current monotherapies also supporting the fight against drug resistance.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3047-3063"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148389990","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":"Artificial Intelligence-Powered Algal Biodiesel: The Future of Biofuel Production Through Data-Driven Biotechnology","authors":"Gyanendra Tripathi","doi":"10.1002/bit.70305","DOIUrl":"10.1002/bit.70305","url":null,"abstract":"<div>\u0000 \u0000 <p>Microalgae are promising feedstocks for sustainable biodiesel production, although their industrial use is currently limited by their biological complexity and high production costs. Recently, artificial intelligence (AI) technologies have been identified as promising, game-changing tools that could optimize algal biodiesel processes by leveraging data-driven biology. The purpose of this Review is to provide an integrated summary of recent developments in machine learning, deep learning, and hybrid models of microalgal biodiesel, analyzing AI frameworks that integrate biological and physical information with microalgal biodiesel process models to improve productivity, biological stability, and sustainability. It is essential to critically analyze the challenges of data quality and explainability in machine learning models, and to explain their use in techno-economic analysis of microalgal biodiesel. In the final part, directions for developing an independent microalgal biorefinery system using artificial intelligence tools are outlined to accelerate the lab-to-production transition of climate-relevant biofuels.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"2968-2983"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148393391","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 and Optimization of a Hollow Fiber-Based Countercurrent Dialysis Process for Continuous Manufacturing of Monoclonal Antibodies","authors":"Vishwanath Hebbi,&nbsp;Mark Brower,&nbsp;Nuno D. S. Pinto","doi":"10.1002/bit.70313","DOIUrl":"10.1002/bit.70313","url":null,"abstract":"<div>\u0000 \u0000 <p>Countercurrent dialysis using hollow fiber modules offers several advantages over both inline diafiltration (ILDF) and hemodialyzer based countercurrent dialysis in terms of buffer requirements and compatibility with GMP manufacturing of biologics. In this work, we evaluate GMP-grade hollow fiber modules of varying lengths (20, 41.5, and 65 cm) and benchmark their performance against an ILDF unit as a reference process for continuous exchange of buffer in manufacturing of monoclonal antibodies. Hollow fiber modules outperformed the ILDF configuration in terms of buffer consumption and pressure build-up, particularly for higher-titer processes. The GMP hollow fiber modules, which are characterized using protein-based molecular weight cut-off (MWCO) specifications, achieved buffer exchange comparable to the reference processes when using endogenous excipients (histidine, methionine) as tracers, while providing a bioprocess-oriented format and documentation suitable for GMP implementation. A D-optimal experimental design was used to define the design space for the hollow fiber countercurrent dialysis step by varying the ratio of dialysate buffer to feed flow (“alpha factor”, 2–5), normalized feed flow rate (0.05–1.5 L·m⁻²·h⁻¹), protein concentration (30–70 mg/mL), and feed conductivity (2–15 mS/cm). The results showed that higher feed concentrations narrowed the acceptable range of normalized feed flow rates and required higher alpha factors to achieve the target buffer exchange. A 5-day continuous run operated at the selected process set-point achieved the desired buffer exchange targets with &gt; 95% product yield, no significant impact on aggregation, and transmembrane pressures below 12 psi (≈80 kPa). Scale-up analysis indicated that the process is suitable for GMP manufacturing at scales up to 3000 L perfusion bioreactors with perfusate titers ranging from 0.6 to 5 mg/mL, while operating within a robust design space.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3118-3133"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148451765","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":"Characterization and Therapeutic Potential of a Novel Lytic Phage-Derived Endolysin PA16cLys Against Uropathogenic Pseudomonas aeruginosa Biofilms","authors":"Shilin Zhang,&nbsp;Han Niu,&nbsp;Yaru Hong,&nbsp;Ruibing Yang,&nbsp;Zahid Shahbaz,&nbsp;Jiansong You,&nbsp;Zhizhong Quan,&nbsp;Jianguang Li,&nbsp;Xiaoyu Li,&nbsp;Yongping Xu,&nbsp;Lili Wang","doi":"10.1002/bit.70301","DOIUrl":"10.1002/bit.70301","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) is a leading cause of urinary tract infections (UTIs). The escalating misuse of antibiotics has led to the emergence of multidrug-resistant strains, necessitating novel therapeutic approaches. Bacteriophages and their encoded lytic enzymes offer unique advantages for antimicrobial therapy and show promising potential for future applications. This study focuses on the characterization of uropathogenic <i>P. aeruginosa</i> phages and their lytic enzymes, investigating their biological properties and potential therapeutic applications. We isolated a novel lytic bacteriophage, vB_PaeS_PA16c (PPA16c), which demonstrated favorable biological characteristics, including a 34% biofilm eradication capacity within 6 h. The phage-encoded endolysin PA16cLys exhibited superior antibacterial efficacy compared to the phage itself. Notably, it achieved significant biofilm reduction (~63%) after 1 h treatment without requiring outer membrane permeabilizers. This endolysin exhibited enhanced antibacterial efficacy in eradicating mature <i>P. aeruginosa</i> biofilms. Regarding host range, PA16cLys displayed broader lytic activity. Both the phage and endolysin demonstrated cross-order lytic activity against other pathogenic bacteria. Crucially, PA16cLys maintained potent inhibitory activity against phage-resistant mutants. These findings provide a theoretical foundation for utilizing phages and lytic enzymes against uropathogenic <i>P. aeruginosa</i> and establish a research basis for developing novel antimicrobial therapy.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3017-3029"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148388747","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":"Purification of Advanced Therapeutics by Flow-Through Pseudo-Affinity Chromatography: Bi-Specific mABs, Fc Fusion Proteins, and Adeno-Associated Viruses","authors":"Wenning Chu,&nbsp;Eduardo Barbieri,&nbsp;Arianna Minzoni,&nbsp;Shriarjun Shastry,&nbsp;Jacob P. LeBarre,&nbsp;Qingbo Shu,&nbsp;Taufika Williams,&nbsp;Xuanwen Li,&nbsp;Divya Chandra,&nbsp;Xuankuo Xu,&nbsp;Jerome Bill,&nbsp;David Roush,&nbsp;Joseph Taris,&nbsp;Patrick Gilbert,&nbsp;Andre Kiesewetter,&nbsp;Oliver Rammo,&nbsp;Michael Schulte,&nbsp;Christopher Major,&nbsp;Trina Mouchahoir,&nbsp;Steven Cramer,&nbsp;Cristiana Boi,&nbsp;Ruben G. Carbonell,&nbsp;Abraham Lenhoff,&nbsp;Stefano Menegatti","doi":"10.1002/bit.70310","DOIUrl":"10.1002/bit.70310","url":null,"abstract":"<p>Modern biopharmaceutical manufacturing requires purification platforms capable of processing structurally and functionally diverse products while addressing the challenge of removing persistent and high-risk host cell proteins (HCPs). Current purification processes utilize adsorbents that operate in bind-and-elute mode, which limits productivity, and often struggle to remove critical impurities. Addressing these limitations, our team developed a flow-through pseudo-affinity chromatography platform that utilizes size-exclusion resins (Monomix Core 500) functionalized with mixed-mode peptide ligands (AviGuard) to remove process-related impurities while maximizing productivity and product integrity. This study evaluates this technology across three therapeutic modalities: two bi-specific monoclonal antibodies (bsAbs) and an Fc-fusion protein from Chinese hamster ovary (CHO) cell culture fluids, and adeno-associated virus serotypes 2 and 9 (AAV2 and AAV9) from HEK293 cell lysates. For the CHO-derived proteins, the platform comprised a pre-capture step with LigaGuard-Monomix Core 500 resin (LGMC500), a Protein A-based capture step, and a polishing step using a single-use size-exclusion mixed-mode (SEMM) resin. This configuration achieved global product yields exceeding 70%, final product pool concentration &gt; 15 mg/mL, monomeric purity ~99%, and excellent cumulative HCP clearance (LRV &gt; 4.5) to residual levels of 60–165 ng/mL (4–11 ppm). For AAV purification, the process utilized a mixed-bed adsorbent comprising dextran-coated charcoal and LGMC500 for removing HCPs, cell metabolites, and media components, followed by a capture step using either CaptureSelect AAVX resin or Natrix CH membranes. The optimization of the charcoal-to-resin volume ratio (1:3) and load volume (~ 10¹⁴ vp per mL of resin) yielded ~50% AAV recovery with a residual HCP level of 350 ng/mL, representing &lt; 100 ng per dose, in compliance with regulatory guidelines. Process productivity ranged from 19 to 23 g/L·hr for CHO-derived proteins and ~7 × 10¹⁴ vp/L·hr for AAV9, positioning this technology in the optimal operating space for modern bioprocessing.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 11","pages":"3079-3105"},"PeriodicalIF":3.9,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70310","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148451710","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}