Biotechnology Journal最新文献

{"title":"Modulation of Antibody Glycosylation in Chinese Hamster Ovary Cells: Experimental and Computational Analyses Provide Mechanistic Insights","authors":"Ranya Pranomphon,&nbsp;Sofia Gialamoidou,&nbsp;Montarop Yamabhai,&nbsp;Ioscani Jiménez del Val,&nbsp;Susan T. Sharfstein","doi":"10.1002/biot.70143","DOIUrl":"10.1002/biot.70143","url":null,"abstract":"<div>\u0000 \u0000 <p>Process conditions and cell culture media components significantly impact glycosylation in ways that are still poorly understood. Here, we evaluated the glycan distribution of CHO-K1 clones producing adalimumab, a biosimilar version of the anti-TNF antibody Humira. To modulate the glycan profile, the cell culture medium was supplemented with manganese and/or galactose. Both manganese and galactose increased galactosylation, and the effects were synergistic. Notably, the levels of Man5 were higher than in Humira, even when galactosylation was similar. qPCR analysis revealed few differences in glycosylation enzyme concentrations between different productivity clones or when 50 µM manganese was added. We modeled the glycosylation pathways using a dynamic mathematical model to elucidate the mechanisms by which high mannose glycans increased and to develop a more predictive approach to culture modulation. Modeling results suggest that the concentration of uridine triphosphate (UTP), a component of activated sugars, is limiting and that increased uridine diphosphate (UDP)-galactose results in decreased UDP-N-acetylglucosamine, limiting complex glycan synthesis.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145372052","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}

{"title":"Overexpression of Matriptase-1 Inhibitor HAI-1 as an Effective Strategy to Reduce Proteolytic Clipping in Chinese Hamster Ovary Cells","authors":"Qinghao Zhang,&nbsp;Fnu Aapjeet,&nbsp;Joyce Chen,&nbsp;Xiaoyan Tang,&nbsp;Gregory F. Pirrone,&nbsp;Alexey A. Makarov,&nbsp;Fengfei Ma,&nbsp;Luke Nelson,&nbsp;Jessica Pan,&nbsp;Ren Liu,&nbsp;Michael J. Hohn,&nbsp;David J. Busch","doi":"10.1002/biot.70141","DOIUrl":"10.1002/biot.70141","url":null,"abstract":"<div>\u0000 \u0000 <p>Proteolytic degradation of recombinant proteins in Chinese hamster ovary (CHO) cells remains a major challenge in biopharmaceutical manufacturing, often reducing product yield and quality. Matriptase-1, a type II transmembrane serine protease, has been identified as a key contributor to unwanted proteolysis. This study investigates hepatocyte growth factor activator inhibitor-1 (HAI-1) overexpression as a strategy to mitigate matriptase-1-mediated degradation in CHO cell culture. Using an IL-12 IgG1 Fc fusion protein and a multi-specific antibody (M molecule) as model proteins, we employed genetic and biochemical approaches to assess the impact of Chinese hamster HAI-1 overexpression on protein quality and yield. Our results demonstrate that HAI-1 overexpression effectively inhibits matriptase-1 activity, achieving up to a 98% reduction in proteolytic clipping while maintaining cell growth, viability, and product quality. Compared to other protease control strategies, HAI-1 overexpression presents a practical and scalable solution that does not disrupt essential cellular functions. These findings establish HAI-1 as a key modulatory tool in CHO-based protein production, with implications for reducing proteolysis of therapeutic proteins and optimizing biopharmaceutical manufacturing processes.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145372030","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}

{"title":"Enhancing Carbon Metabolic Flow Ability for Titer Improvement of L-Lactic Acid in Bacillus coagulans Through Heavy Ion Mutagenesis","authors":"Weibin Ma,&nbsp;Wenwen Shan,&nbsp;Jihong Chen,&nbsp;Zhen Zhang,&nbsp;Wei Hu","doi":"10.1002/biot.70142","DOIUrl":"10.1002/biot.70142","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Bacillus coagulans</i> is a preferred producer for lactic acid, still facing the challenge of poor carbon metabolic flow ability. Here, an engineered <i>B. coagulans</i> strain P14 with improved carbon metabolic flow ability was developed for the first time through heavy ion mutagenesis combined with an enrichment strategy of acetic acid and sodium acetate, showing increased titer, productivity and yield of L-lactic acid compared to the parental strain. Notably, the activities of four enzymes involved in the glycolytic pathway of <i>B. coagulans</i> P14 were also higher. For batch fermentations, the engineered <i>B. coagulans</i> P14 strain showed improved production economy than the parental strain, which can produce 124.6 g/L L-lactic acid with a productivity of 7.1 g/L/h and a yield of 93% in a 5-L bioreactor under unsterile conditions, showing an increase of 6.0%, 24.3%, and 8.1% compared to the parental strain, respectively. This work showcases a method for rapidly breeding a robust lactic acid-producing strain through heavy ion mutagenesis combined with an enrichment strategy.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145372027","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}

{"title":"mRNA Stability: An Unresolved Challenge for Broad Therapeutic Applications","authors":"Yiming Wang,&nbsp;Xiaoxue Wang,&nbsp;Yuan Lu","doi":"10.1002/biot.70146","DOIUrl":"10.1002/biot.70146","url":null,"abstract":"<div>\u0000 \u0000 <p>From infectious diseases and cancers to various rare diseases, mRNAs have demonstrated considerable therapeutic potential for a wide range of diseases. However, due to their single-stranded structure, mRNA molecules are vulnerable to enzyme-mediated degradation. Therefore, the inherent instability of mRNA poses a significant challenge. In this review, we explore strategies to slow down the degradation rate, such as removing degradative enzymes, adding protective substances, and optimizing storage and transport conditions to enhance mRNA stability. Furthermore, optimizing the sequence and structure of mRNAs is crucial for improving stability, which can be significantly aided by fine-tuning the sequences of the 5' untranslated region, open reading frame, and 3' untranslated region, along with introducing various RNA modifications. The design of novel mRNA structures, including circular mRNA and self-amplifying RNA, also offers novel approaches for enhancing mRNA stability. Additionally, we briefly introduce the use of mRNA delivery materials for improving stability and discuss current challenges and future directions in mRNA development. With ongoing technological advancements and the gradual maturation of the market, mRNA is set to play an increasingly significant role in versatile biotechnology fields.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145371990","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}

{"title":"Efficient Agrobacterium-Mediated Transformation of Green Arabidopsis Suspension Cells","authors":"Matthias Buntru,&nbsp;Stefano Di Fiore,&nbsp;Nils Hahnengress,&nbsp;Helga Schinkel,&nbsp;Stefan Schillberg,&nbsp;Greta Nölke","doi":"10.1002/biot.70145","DOIUrl":"10.1002/biot.70145","url":null,"abstract":"<p>Photosynthetic plant cell suspension cultures are a valuable experimental system for analyzing various physiological processes. This system bypasses the structural complexity of the whole plant organism and can be manipulated under uniform conditions. However, achieving a highly efficient and consistent transformation of plant suspension cells remains challenging. By using green fluorescent protein (GFP) and a microplate confocal imaging system for high-throughput analysis, we optimized the transformation of green <i>Arabidopsis</i> suspension cells to infect almost 100% of the cells. Key elements of our protocol included using the hypervirulent <i>Agrobacterium tumefaciens</i> strain AGL1, co-cultivating agrobacteria and suspension cells on solidified medium plates, and adding AB minimal salts and the surfactant Pluronic F68. The presented method can significantly increase the transformation rate of plant suspension cells, facilitating the introduction of genetic pathways for producing industrial, cosmetic, or pharmaceutical compounds in these systems.</p>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70145","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145353125","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}

{"title":"A CFD-Based Digital Framework for Scaling Optimization of Orbital Rocking Bioreactors","authors":"Soo Hyun Ryu,&nbsp;Young Jin Kim,&nbsp;Jae Hong Jeon,&nbsp;Minjun Ji,&nbsp;Tae Hyeong Kim,&nbsp;Kyung Nam Kim,&nbsp;Dong-Hoon Yang,&nbsp;Tack-Joong Kim,&nbsp;Jong Kwang Hong","doi":"10.1002/biot.70138","DOIUrl":"10.1002/biot.70138","url":null,"abstract":"<div>\u0000 \u0000 <p>Robust scale-up of bioreactor systems requires hydrodynamic similarity across scales to ensure consistent cell culture performance. This study presents a computational fluid dynamics (CFD)-guided digital framework for optimizing the scale-up of a novel orbital rocking bioreactor, CELBIC, from CELBIC5 (working volume 1–2 L) to CELBIC50 (working volume 10–20 L). Using lattice Boltzmann-based simulations, key hydrodynamic parameters, including velocity, shear stress, and energy dissipation rate, were evaluated across various working volumes, inclination angles, and agitation speeds. Compared with conventional scale-up criteria, such as average P/V, we propose a comprehensive and digitalized scale-up optimization framework: a root mean square error (RMSE)-based method comparing the full spatial and temporal distributions of various CFD variables to quantify similarities among scaled-up conditions. This approach allows identification of the best-matching scaled-up condition and further refinement using response surface analysis. The optimized condition (10 L, 7°, and 19 rpm) exhibited the lowest combined RMSE relative to the reference condition (CELBIC5 at 1 L, 6°, and 30 rpm), suggesting substantial hydrodynamic equivalence. Overall, this study demonstrates how integrating CFD- and RMSE-based analyses enables rational scale-up of orbital rocking bioreactors, offering a systematic strategy for digital process development in single-use systems.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336072","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}

{"title":"Immune Response of Zika prM Peptide PEG Conjugates in Peripheral Blood Mononuclear Cells","authors":"Yogita Gupta,&nbsp;Manoj Baranwal,&nbsp;Bhupendra Chudasama","doi":"10.1002/biot.70140","DOIUrl":"10.1002/biot.70140","url":null,"abstract":"<div>\u0000 \u0000 <p>Despite the challenge in generating adequate immunogenicity, peptide-based vaccines offer a significant benefit in terms of improving immunity when combined with conjugates. In previous studies, three computationally designed immunogenic peptides of the Zika prM protein were reported to induce an immune response. In the current work, reported peptides were conjugated with polymer, polyethylene glycol 400, with the intention of enhancing the immune response. Conjugation was confirmed by Fourier transform infrared spectroscopy. We reckoned the immune response of all three peptide-PEG conjugates (MPC1, MPC2, and MPC3) on 10 different healthy blood samples. Assessment of MPC-induced human peripheral blood mononuclear cell (PBMC) proliferation and interferon-gamma (IFN-γ) secretion was done with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and a sandwich enzyme-linked immunosorbent assay (ELISA). Conjugate MPC1 showed significantly enhanced cell proliferation in five and MPC2 and MPC3 in six samples, compared to the peptides. In case of IFN-γ release, conjugate MPC1 exhibited significant results in six, MPC2 in five, and MPC3 in seven samples elevated results, in contrast to peptides alone. Thus, conjugation of PEG to immunogenic peptides could be an effective way to increase the peptide immunogenicity, although further experimental validations are required before considering them as candidates for a vaccine against Zika virus infection.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336075","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}

{"title":"Correction to “Enhanced Growth of Bone Marrow-Derived Mesenchymal Stem Cells on the Microcarriers Tethered With Engineered Basic Fibroblast Growth Factor”","authors":"","doi":"10.1002/biot.70139","DOIUrl":"10.1002/biot.70139","url":null,"abstract":"<p>S. Pasanen, A. Mootha, I. Hirata, K. Tanimoto, and K. Kato, “Enhanced Growth of Bone Marrow-Derived Mesenchymal Stem Cells on the Microcarriers Tethered With Engineered Basic Fibroblast Growth Factor,”<i> Biotechnology Journal</i> 20 (2025): e70057.</p><p>In Figure 5 (d), a labeling inserted in the graph “[CBP1-bFGF]” was incorrect. This should have read “[CBP2-bFGF]”.</p><p>We apologize for this error.</p>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70139","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145278454","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}

{"title":"Development of Microvascular Network in Microfluidic Brain-on-a-Chip Models In Vitro: A Multidisciplinary Review","authors":"Alla B. Salmina,&nbsp;Mikis R. Saridis,&nbsp;Vitaly V. Ryzhkov,&nbsp;Sofia A. Korsakova,&nbsp;Anton S. Averchuk,&nbsp;Daniil A. Bystrov,&nbsp;Anastasiia A. Barbasheva,&nbsp;Pavel A. Libet,&nbsp;Alexei K. Kuguk,&nbsp;Leonid Y. Polynkin,&nbsp;Lyubov N. Muravyova,&nbsp;Kirill A. Mamaev,&nbsp;Ruslan Sh. Alibekov,&nbsp;Ivan A. Kushnir,&nbsp;Egor V. Yakovlev,&nbsp;Ilya A. Rodionov,&nbsp;Stanislav O. Yurchenko","doi":"10.1002/biot.70126","DOIUrl":"https://doi.org/10.1002/biot.70126","url":null,"abstract":"<div>\u0000 \u0000 <p>Microfluidic brain-on-a-chip and angiogenesis-on-a-chip models have been employed by leading research groups around the world. These models have great potential, but they have not yet been used in the context of personalized medicine and diagnostics, nor have they been widely adopted for testing drug candidates. The reproduction of a physiologically relevant in vitro brain-on-a-chip model with essential similarity to in vivo structural and functional characteristics, including integrity and complexity of the brain tissue, limited and controlled permeability of barriers, and plastic changes induced by brain stimulation or injury, is a highly challenging task that requires a comprehensive approach and a tour de force in advanced biomedical engineering and living soft matter physics. This approach necessitates not only the combination of well-established chip production technologies and cell biology protocols but also the integration of knowledge from neuroscience, (bio)physics, hydrodynamics, material science, (bio)chemistry, electronics, and soft matter physics. This review covers current understandings of the establishment of microvascular network formation in the active brain in vivo, as well as analysis of novel approaches to reconstruct the basic mechanisms of cerebral angiogenesis and barrier genesis in a physiologically relevant brain-on-a-chip model. The novel concept, which employs the application of an external electric field to stimulate vasculogenesis/angiogenesis on a chip, demonstrates how a synergistic approach may help to solve this non-trivial problem and to establish the vascularized brain tissue sensitive to various stimuli and suitable for the high-precision analysis of its functional activity and plasticity.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145272501","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}

{"title":"Low Concentration Ionic Liquid-Assisted Efficient Enzymatic Degradation of Xanthan","authors":"Chunshu Chen,&nbsp;Fan Yang,&nbsp;Le Zhong,&nbsp;Jinyun Gu,&nbsp;Weiming Liu,&nbsp;Wenbo Li,&nbsp;Ruiyu Shen,&nbsp;Zhimin Yu","doi":"10.1002/biot.70136","DOIUrl":"10.1002/biot.70136","url":null,"abstract":"<div>\u0000 \u0000 <p>One of the main obstacles to preparing oligoxanthan through the enzymatic degradation of xanthan gum (XG) is its dense structure. To achieve the large-scale production of oligoxanthan, it is necessary to perturbate the conformation of XG and increase enzyme accessibility. This research developed an efficient system for the enzymatic degradation of XG assisted by low concentrations of ionic liquids (ILs). Structural characterization results indicated that a low concentration of [EMIM]DEP (1%, v/v) not only loosens the structure of XG but also alters the secondary structure of enzyme, thereby dramatically increasing its hydrolytic activity and eventually enhancing the production of oligoxanthan. The results were further elucidated by molecular dynamics (MD) simulation. Under the hydrolysis conditions of 1.5 mg/mL <i>Aspergillus niger</i> cellulase (<i>An</i>Cel), 0.2 mg/mL XG, and 1% (v/v) [EMIM]DEP at 40°C and pH 5.0 for 20 min, XG can be hydrolyzed into a mixture with a glucose equivalent of 32.25%. The degree of polymerization (DP) of XG hydrolysates was 3–10. Moreover, the xanthan hydrolysates obtained using this system exhibit excellent antioxidant activity. This study proposed an effective method for the preparation of functional oligosaccharides, which is of significant importance for the valorization of XG.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 10","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145256912","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}