Biotechnology Journal最新文献_第8页

Engineering Design of Artificial Phase-Separating Proteins 人工相分离蛋白质的工程设计。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70246

Yongjun Mao, Lu Bao, Gaoshuai Li, Qicheng Zhu, Yanchao Han, Xudong Wang

{"title":"Engineering Design of Artificial Phase-Separating Proteins","authors":"Yongjun Mao, Lu Bao, Gaoshuai Li, Qicheng Zhu, Yanchao Han, Xudong Wang","doi":"10.1002/biot.70246","DOIUrl":"10.1002/biot.70246","url":null,"abstract":"<div>\u0000 \u0000 Phase-separating proteins are evolutionarily conserved biomolecules that orchestrate key cellular processes. Although the biophysical principles underlying liquid–liquid phase separation (LLPS)—including multivalent interactions, intrinsically disordered regions (IDRs), and the sticker–spacer framework—are now well established, translating these insights into actionable engineering guidelines remains open challenges. Here, we present a design-oriented synthesis for engineering artificial phase-separating proteins. Drawing on a curated dataset, we characterize the physicochemical patterns associated with LLPS behavior, we evaluate current computational predictors, revealing systematic blind spots in hydrophobicity-rich or modular repeat architectures. Building on these insights, we propose modular design heuristics that encompasses sequence-level composition and patterning, valency control via repeat architecture, and topological fusion strategies including terminal tags, telechelic constructs, and interleaved block architectures. These design levers are mapped onto applications such as protein purification, catalytic condensates, stimuli-responsive systems, and mechanically hydrogels and fibers. Finally, we present an integrated design–build–test–learn (DBTL) framework that bridges predictive modeling, experimental iteration, and functional validation. By treating phase separation as an engineerable rather than merely emergent property, this review offers both a conceptual map and a practical roadmap for the rational construction of artificial phase-separating proteins across biotechnological and biomedical contexts.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148040478","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}

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IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28

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IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28

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Nanoparticle-Based Radiosensitization in Breast Cancer: A Multimodal Approach to Therapy and Immune Regulation 基于纳米粒子的乳腺癌放射增敏：治疗和免疫调节的多模式方法。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70242

Saleh A. S. AlAbdulhadi, Ali A. Majeed, Djamila Polatova, Muath Suliman, G. PadmaPriya, Karthikeyan Jayabalan, Vimal Arora, Priya Priyadarshini Nayak, REYADH R AL-RASHIDI, Wesam R. Kadhum

{"title":"Nanoparticle-Based Radiosensitization in Breast Cancer: A Multimodal Approach to Therapy and Immune Regulation","authors":"Saleh A. S. AlAbdulhadi, Ali A. Majeed, Djamila Polatova, Muath Suliman, G. PadmaPriya, Karthikeyan Jayabalan, Vimal Arora, Priya Priyadarshini Nayak, REYADH R AL-RASHIDI, Wesam R. Kadhum","doi":"10.1002/biot.70242","DOIUrl":"10.1002/biot.70242","url":null,"abstract":"<div>\u0000 \u0000 Radiotherapy is essential for breast cancer treatment, especially after lumpectomy. Its effectiveness, however, is hindered by tumor resistance, which causes recurrence and poorer outcomes. In the last decade, nanoparticle-mediated radiosensitization has emerged as a promising strategy to improve radiotherapy in breast cancer. Metal nanoparticles, such as gold, silver, platinum, and bismuth, increase the localized radiation dose through secondary electron emission and generate reactive oxygen species (ROS). Polymeric, liposomal, and hydrogel nanocarriers deliver radiosensitizers directly to tumors with sustained release. These features increase cytotoxicity and reduce hypoxia-related radioresistance. Nanoparticles also inhibit DNA repair pathways and modulate the tumor microenvironment, which enhances the radiation response. Additionally, nanoparticles can boost the effects of immunotherapy by activating dendritic cells, strengthening immune checkpoint blockade, and promoting anti-tumor immune memory. Recent preclinical studies have shown that combining nanoparticles loaded with natural compounds, such as curcumin, polyphenols, flavonoids, and metformin, increases oxidative stress and radiosensitivity in breast tumors while sparing healthy tissue. We provide an overview of nanoparticle-mediated radiosensitization mechanisms and novel supporting approaches for achieving better tumor responses with reduced toxicity.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148040438","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}

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Issue Information: Biotechnology Journal 6/2026 期刊信息：Biotechnology Journal 6/2026

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70254

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IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28

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Engineering Secretory (S)IgA to Facilitate Affinity Purification for Large Scale Downstream Processing 促进大规模下游加工亲和纯化的工程秘书处(S)IgA。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70245

Tim H. Szeto, Julian K-C. Ma

{"title":"Engineering Secretory (S)IgA to Facilitate Affinity Purification for Large Scale Downstream Processing","authors":"Tim H. Szeto, Julian K-C. Ma","doi":"10.1002/biot.70245","DOIUrl":"10.1002/biot.70245","url":null,"abstract":"Secretory (S)IgA class antibodies play a vital role in first line immunological defence at mucosal surfaces. Passive immunisation with monoclonal SIgA antibodies could offer a completely novel application for a biologics market currently dominated by systemic IgG class monoclonal antibodies. Recent advances have made large scale manufacture of SIgA mAbs feasible, but downstream affinity purification remains a hurdle because IgA antibodies usually have poor affinity for Protein A, the most widely used affinity ligand for antibodies. This research addresses this problem by engineering chimaeric secretory component (SC), containing IgG Fc domains (CH2─CH3) which confer Protein A binding. SC is a component of SIgA and consists of five immunoglobulin-like domains. We demonstrate that expressing recombinant monoclonal SIgA with SC domains D2–D3 or D3–D4 replaced by IgG Fc domains (CH2─CH3) enables Protein A affinity purification of SIgA, without affecting antibody assembly or antigen binding functionality. Inserting the Protein A binding site into SC, results in purification of SIgA and free SC, which could then be separated according to size, but importantly not SIgA assembly intermediates such as dimeric IgA, that has a similar molecular size to SIgA. The use of SC-Fc fusions could streamline SIgA mAb purification and unlock their clinical potential.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70245","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148040444","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

High-Yield Production and Cost-Effective Purification of an Alkali-Resistant Hexameric Protein A Variant for Antibody Affinity Chromatography 抗碱六聚体蛋白A变体抗体亲和层析的高效生产和低成本纯化。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70247

Carolina Mota Natal, Claudia Lacombe, Alexander Zollner, Catarina Domingos, José Mendes, André Nascimento, Ana Margarida Gonçalves Carvalho Dias, Arménio Jorge Moura Barbosa, Cristina Peixoto, Birgit Wiltschi, Alois Jungbauer, Ana Cecília Afonso Roque

{"title":"High-Yield Production and Cost-Effective Purification of an Alkali-Resistant Hexameric Protein A Variant for Antibody Affinity Chromatography","authors":"Carolina Mota Natal, Claudia Lacombe, Alexander Zollner, Catarina Domingos, José Mendes, André Nascimento, Ana Margarida Gonçalves Carvalho Dias, Arménio Jorge Moura Barbosa, Cristina Peixoto, Birgit Wiltschi, Alois Jungbauer, Ana Cecília Afonso Roque","doi":"10.1002/biot.70247","DOIUrl":"10.1002/biot.70247","url":null,"abstract":"Recombinant Staphylococcal Protein A is the most widely used affinity ligand for industrial antibody purification, as well as for research and discovery of new antibodies. It has become a platform technology throughout the antibody manufacturing industry for the direct capture of products from a clarified cell culture supernatant. However, protein A adsorbents contribute significantly to manufacturing costs in the early stage of process development. In this work, we aimed to increase the production yield of an engineered protein A ligand comprising six repeats of the alkali-resistant protein A-derived Z domain, with a terminal cysteine residue for adsorbent immobilization (Z6AlkC). We coupled the extracellular production of Z6AlkC via secretion in high-cell-density, carbon-limited bioreactor cultures, with a simple and fast ultradiafiltration step for purification. Our bioprocess achieved 4 g/L of secreted Z6AlkC with an 89% purity, which is an improvement from previous reports that obtained intracellular expression and lower yield. We further purified the Z6AlkC protein by IgG-based affinity chromatography (Z6AlkC-P) and ultradiafiltration (Z6AlkC-F) and used it as an affinity ligand for antibody purification, as a proof of functionality. Both ligands yielded similar purification antibody behavior, showing that high-performance protein A-based ligands can be produced in scalable, high-productivity, and cost-effective methods.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 6","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70247","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148040447","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

Low-Shear Circulation and Layered Regeneration: Coupled Analysis of RCCS-CFD for In Vitro Construction of Active Osteochondral Tissues 低剪切循环与层状再生：体外构建活性骨软骨组织的RCCS-CFD耦合分析。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70248

Weiting Guo, Hezhi Chen, Xueyan Hu, Jingjing Zhu, Xinyue Liu, Yanqiu Pan, Yuen Yee Cheng, Yue Kang, Xiangqin Li, Jiaquan Zhao, Kedong Song

{"title":"Low-Shear Circulation and Layered Regeneration: Coupled Analysis of RCCS-CFD for In Vitro Construction of Active Osteochondral Tissues","authors":"Weiting Guo, Hezhi Chen, Xueyan Hu, Jingjing Zhu, Xinyue Liu, Yanqiu Pan, Yuen Yee Cheng, Yue Kang, Xiangqin Li, Jiaquan Zhao, Kedong Song","doi":"10.1002/biot.70248","DOIUrl":"10.1002/biot.70248","url":null,"abstract":"<div>\u0000 \u0000 The rotating cell culture system (RCCS) is widely used in 3D in vitro culture due to its ability to provide a low shear environment and optimal mass transfer. However, current understanding of the shear stress and fluid flow distribution within the RCCS remains limited. In this study, a dynamic culture environment of RCCS was constructed in vitro. The three-dimensional spatial structure of chitosan7/gelatin3/Nano-hydroxyapatite (Cs7/Gel3/nHAP) multilayer composite scaffold constructs and bone-derived scaffold structures was used to guide the directional differentiation of human adipose-derived stem cells (ADSCs). Then, computational fluid dynamics was used to simulate and predict the stress and fluid flow distribution of ADSCs-Cs7/Gel3/nHAP constructs and ADSCs-derived-bone scaffold constructs under different rotational speeds in RCCS. It was confirmed that the flow field environment in RCCS was a relatively stable annular laminar flow. Moreover, during the cultivation process in RCCS, the complex was subjected to a uniformly distributed shear force. Finally, the optimal operating conditions of RCCS were selected when two kinds of cell-scaffold bone constructs were placed in RCCS, and it has been experimentally verified that culturing bionic osteochondral scaffolds in the RCCS demonstrates significantly higher ADSCs proliferative activity and differentiation marker expression compared to static culture conditions.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148040499","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 Recombinant Adeno-Associated Virus (rAAV) Biomanufacturing: Design of Experiment (DOE) Enabled Transfection Optimization for Maximum Full Capsid Yield and Robust Scale-Up 增强重组腺相关病毒（rAAV）生物制造：实验设计（DOE）使转染优化，以获得最大的全衣壳产量和稳健的放大。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-28 DOI: 10.1002/biot.70249

Alexandra Bogdanovic, Nicholas Donohue, Brian Glennon, Susan Mc Donnell, Jessica Whelan

{"title":"Enhanced Recombinant Adeno-Associated Virus (rAAV) Biomanufacturing: Design of Experiment (DOE) Enabled Transfection Optimization for Maximum Full Capsid Yield and Robust Scale-Up","authors":"Alexandra Bogdanovic, Nicholas Donohue, Brian Glennon, Susan Mc Donnell, Jessica Whelan","doi":"10.1002/biot.70249","DOIUrl":"10.1002/biot.70249","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) gene therapies are a promising class of therapeutics. While their production via triple transfection in a single cell line offers flexibility regarding modification of the transgene, it faces significant challenges, particularly regarding predominance of empty capsids, reproducibility of process performance (titre and product quality), and successful scale-up. This study addresses these limitations by employing a two-stage Design of Experiment (DOE) approach to optimize triple transfection for rAAV5 viral vector production. An initial screening design systematically evaluated a comprehensive set of factors for their impact on transfection efficiency, genome titre, capsid titre, and the ratio of full to empty capsids. Four highly influential factors identified during screening were further investigated in a second-stage response surface design. Statistical analysis confirmed that DNA amount, complexation time, FectoVir-AAV volume, and the ratio of pTransgene were the most critical determinants of performance. The optimal conditions were successfully scaled up from 6 well plates to a 50 L Wave reactor. Coupled with the implementation of an alternative harvest strategy, transfection efficiencies, capsid, and genome titre were maintained while there was an increase in % full capsids to 73% at scale demonstrating a viable, robust, and scalable manufacturing strategy for high-quality rAAV5 vectors.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 6","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70249","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148040422","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