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Adaptabodies: Retargeting a Predefined Monoclonal Antibody With Bispecific Nanobodies for Antibody Therapy 适应体:用双特异性纳米体重新靶向预定义的单克隆抗体用于抗体治疗
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-19 DOI: 10.1002/biot.70089
Andrés González-Techera, Gabriel Lassabe, José A. Chabalgoity, Cecilia Vallejo Garín, Julio Guarnaschelli, Gualberto González-Sapienza

Monoclonal antibody (MoAb) therapy is a cornerstone in treating cancer, inflammatory diseases, and infections. However, the development of new monoclonal antibodies is labor-intensive, costly, and species-specific, limiting their accessibility in veterinary medicine and slowing innovation in human therapies. In this work, we introduce adaptabodies, bispecific nanobody constructs that repurpose existing MoAbs, of irrelevant specificity, by bridging their idiotype to a new antigen. As a proof of concept, we tested this strategy using a model murine MoAb combined with a neutralizing adaptabody that redirected its binding to tetanus toxin and resulted in mice surviving at least for 8 days when challenged with 1 lethal dose 100 (LD100). Mice challenged with 20 LD100 of tetanus toxin and treated with two neutralizing adaptabodies survived 8 days and increasing five-fold the amount of the MoAb increased survival to at least 20 days. This approach reduces the need to develop new monoclonal antibodies for each disease, requiring only the retargeting domain to be generated. Moreover, a single MoAb could be developed for species lacking therapeutic MoAbs for adaptabody-mediated treatment. The increasing approval of monoclonal antibodies for treatment of both humans and companion animals underscores the relevance of this flexible and scalable strategy.

单克隆抗体(MoAb)疗法是治疗癌症、炎症性疾病和感染的基石。然而,新的单克隆抗体的开发是劳动密集型的,昂贵的,并且是物种特异性的,限制了它们在兽药中的可及性,减缓了人类治疗的创新。在这项工作中,我们引入了适应体,双特异性纳米体结构,通过桥接其独特型到新的抗原,重新利用现有的不相关特异性的MoAbs。为了验证这一概念,我们使用了一种模型小鼠MoAb结合了一种中和性适应体来测试这一策略,这种适应体可以将其与破伤风毒素的结合重新定向,并导致小鼠在1致死剂量100 (LD100)的攻击下存活至少8天。用20 LD100的破伤风毒素攻毒和两种中和性适应体处理的小鼠存活8天,增加5倍的摩阿布量使小鼠存活至少20天。这种方法减少了为每种疾病开发新的单克隆抗体的需要,只需要生成重靶向结构域。此外,对于缺乏治疗性MoAb的物种,可以开发单一的MoAb用于适应体介导的治疗。越来越多的单克隆抗体被批准用于治疗人类和伴侣动物,强调了这种灵活和可扩展策略的相关性。
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引用次数: 0
Exploring Plant α-Amylase Inhibitors: Mechanisms and Potential Application for Insect Pest Control 探索植物α-淀粉酶抑制剂:防治害虫的机制和潜在应用
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-19 DOI: 10.1002/biot.70098
Marcos Fernando Basso, Arnubio Valencia-Jiménez, Fabrizio Lo Celso, Isabel Rodrigues Gerhardt, Thomas Joseph V. Higgins, Maria Fatima Grossi-de-Sa

α-Amylases are found in microbes, plants, and animals, including insect pests. They play crucial roles in catalyzing the hydrolysis of α-1,4-glucan bonds within starch, glycogen, and related carbohydrates, forming shorter oligomers. In green plants, these enzymes are pivotal for starch degradation during photosynthesis and seed germination, whereas in phytophagous insect pests, they predominantly facilitate seed parasitism by degrading raw starch granules. Amylase inhibitors in plants appear to function as part of their defense against pests and pathogens. In the context of insect pests, some of these amylase inhibitors can target α-amylases in the digestive system of certain insects. Both mono- and dicotyledonous plants harbor multiple genes encoding proteinaceous α-amylase inhibitors. Previous studies have demonstrated that α-amylase inhibitors, whether produced in vitro or overexpressed in transgenic plants, can exhibit entomotoxic activity against certain insect pests. Field trials involving transgenic plants that overexpress α-amylase inhibitors have been conducted, laying the foundation for the potential commercialization of crops engineered with these genes. Herein, this review explores the molecular interactions between plant α-amylase inhibitors and insect α-amylases, shedding light on the underlying mechanisms of action, structural diversity, and assessing the broader biotechnological applications of this promising strategy.

α-淀粉酶存在于微生物、植物和动物,包括害虫中。它们在催化淀粉、糖原和相关碳水化合物中的α-1,4-葡聚糖键水解,形成更短的低聚物方面起着至关重要的作用。在绿色植物中,这些酶在光合作用和种子萌发过程中是淀粉降解的关键,而在植食害虫中,它们主要通过降解原料淀粉颗粒来促进种子寄生。植物中的淀粉酶抑制剂似乎是它们防御害虫和病原体的一部分。在害虫的背景下,这些淀粉酶抑制剂中的一些可以靶向某些昆虫消化系统中的α-淀粉酶。单子叶植物和双子叶植物都含有多个编码蛋白α-淀粉酶抑制剂的基因。先前的研究表明,α-淀粉酶抑制剂,无论是体外产生的还是在转基因植物中过表达的,都能对某些害虫表现出虫毒活性。已经进行了涉及过表达α-淀粉酶抑制剂的转基因植物的田间试验,为用这些基因改造的作物的潜在商业化奠定了基础。本文综述了植物α-淀粉酶抑制剂与昆虫α-淀粉酶之间的分子相互作用,揭示了其潜在的作用机制、结构多样性,并评估了这一有前景的策略的更广泛的生物技术应用。
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引用次数: 0
Uncovering Growth-Inhibitory Metabolic Byproducts in HEK293 Fed-Batch Cultures and Strategies for Their Control HEK293补料批培养中生长抑制代谢副产物的发现及其控制策略
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-19 DOI: 10.1002/biot.70087
Cameron Harrington, Bhanu Chandra Mulukutla

Human Embryonic Kidney 293 (HEK293) cells are currently one of the preferred host cell lines for the production of biologics, specifically, AAV-based viral vectors. These fast-growing cells consume significant amounts of nutrients and often convert them into byproducts such as lactate and ammonia. In fed-batch cultures, accumulation of lactate and ammonia to high levels can inhibit cell proliferation. In this study, we demonstrate that lactate and ammonia accumulation alone doesn't fully explain the growth inhibition observed in HEK293 fed-batch cultures. Growth inhibition was noted even when the residual levels of these byproducts were well controlled. Instead, we show that several previously unknown compounds accumulate in HEK293 cell fed-batch cultures, some of which can inhibit HEK293 cell growth either individually or synergistically. Many of these newly identified compounds are intermediates or byproducts of amino acid catabolism. When residual levels of the source amino acids for these novel byproducts were controlled in the low concentration range (∼1 mM) in HEK293 fed-batch cultures, lactate accumulated to higher levels, causing growth inhibition. This prompted the use of High-end pH Delivery of Glucose (HIPDOG), a control strategy that limits lactate production by keeping low residual concentrations of glucose. In HIPDOG cultures, controlling the source amino acids at low concentrations resulted in lower accumulations of the corresponding growth-inhibitory byproducts when compared to the control HIPDOG conditions with typical levels of amino acids. This led to higher viable cell densities (VCD) and viabilities in low amino acid conditions. Strategies that reduce byproduct accumulation, whether classical or novel byproducts, in HEK293 fed-batch processes can result in enhanced VCDs, potentially leading to higher volumetric productivities.

人胚胎肾293 (HEK293)细胞目前是生产生物制品,特别是基于aav的病毒载体的首选宿主细胞系之一。这些快速生长的细胞消耗大量的营养物质,并经常将其转化为副产品,如乳酸盐和氨。在饲料分批培养中,乳酸和氨积累到较高水平可以抑制细胞增殖。在这项研究中,我们证明了乳酸和氨的积累并不能完全解释HEK293分批饲养培养中观察到的生长抑制。即使这些副产品的残留水平得到很好的控制,也会注意到生长抑制。相反,我们发现几种以前未知的化合物在HEK293细胞补料分批培养中积累,其中一些化合物可以单独或协同抑制HEK293细胞的生长。这些新发现的化合物中有许多是氨基酸分解代谢的中间产物或副产物。当这些新型副产物的源氨基酸残留水平在HEK293补料批培养物中控制在低浓度范围(~ 1 mM)时,乳酸积累到较高水平,导致生长抑制。这促使人们使用高端pH递送葡萄糖(HIPDOG),这是一种通过保持低残留葡萄糖浓度来限制乳酸生成的控制策略。在HIPDOG培养中,与具有典型氨基酸水平的HIPDOG对照条件相比,在低浓度下控制源氨基酸导致相应的生长抑制副产物的积累较低。这导致较高的活细胞密度(VCD)和在低氨基酸条件下的存活率。在HEK293进料批工艺中,减少副产物积累的策略,无论是传统的还是新的副产物,都可以导致vcd的增强,从而可能导致更高的体积生产率。
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引用次数: 0
A Miniaturized Centrifugal Pump ECMO System Enhances Hemocompatibility in Small Animal Models 小型离心泵ECMO系统提高小动物模型的血液相容性
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-17 DOI: 10.1002/biot.70103
Zhen Yang, Yiai Li, Jingyi Peng, Danhe Jia, Youpeng Zhang, Wenxing Huo, Zhigang Liu, Xian Huang

Extracorporeal membrane oxygenation (ECMO) is a fundamental treatment for cardiovascular and severe pulmonary diseases, with small animal models providing critical insights into organ protection during cardiopulmonary bypass and ECMO therapy. Conventional roller pumps, however, induce hemolysis and organ injury through repetitive compression-shear exposure, severely limiting their utility. While centrifugal pumps reduce shear stress and are effective in large animal ECMO, their flow range and priming volume are unsuitable for small animals. Here, we present a miniaturized, fully integrated ECMO system with an optimized centrifugal pump tailored for small animal models. The system reduces shear stress, minimizes blood damage, and enhances organ protection. Integrated multi-parameter sensors enable real-time monitoring of blood flow, pressure, and temperature, thereby streamlining setup and improving perioperative support. In a 6-h experiment, the system demonstrated significant hemolysis suppression, improved renal function (with reduced levels of Neutrophil Gelatinase-Associated Lipocalin, or NGAL), and stable hemodynamics. This innovation offers safer extracorporeal support for small animal studies on cardiovascular diseases, organ recovery, and ECMO mechanisms, furthering research into therapeutic interventions. By addressing key limitations of existing pumps, the system provides a reliable platform for exploring hemodynamic and pathophysiological processes in ECMO treatment, establishing a foundation for future preclinical investigations.

体外膜氧合(ECMO)是心血管和严重肺部疾病的基本治疗方法,小动物模型为体外循环和ECMO治疗期间的器官保护提供了重要见解。然而,传统的滚柱泵通过反复的压缩-剪切暴露诱导溶血和器官损伤,严重限制了它们的应用。离心泵可以降低剪切应力,在大型动物ECMO中是有效的,但其流量范围和启动体积不适用于小动物。在这里,我们提出了一个小型化,完全集成ECMO系统与一个优化的离心泵量身定制的小动物模型。该系统减少剪切应力,减少血液损伤,增强器官保护。集成的多参数传感器能够实时监测血流、压力和温度,从而简化设置并改善围手术期支持。在6小时的实验中,该系统显示出明显的溶血抑制,肾功能改善(中性粒细胞明胶酶相关脂钙蛋白或NGAL水平降低)和稳定的血流动力学。这一创新为心血管疾病、器官恢复和ECMO机制的小动物研究提供了更安全的体外支持,进一步研究治疗干预措施。通过解决现有泵的主要局限性,该系统为探索ECMO治疗中的血流动力学和病理生理过程提供了可靠的平台,为未来的临床前研究奠定了基础。
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引用次数: 0
Evaluation of “Difficult-to-Express” Monoclonal Antibodies in a CHO-Based Hybrid Site-Specific Integration System Under Industrially Relevant Conditions 在工业相关条件下,基于cho的杂交位点特异性整合系统中“难以表达”单克隆抗体的评价
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-17 DOI: 10.1002/biot.70102
Alana C. Szkodny, Kelvin H. Lee

Variation in the primary sequence of monoclonal antibodies (mAbs) can negatively affect their behavior in biopharmaceutical manufacturing platforms, and efforts to identify mAbs with poor “developability” characteristics lack robust methods for assessing mAb expression from an industrially relevant platform. Recent advancements in site-specific integration-based (SSI) platforms in Chinese hamster ovary (CHO) cells can mitigate the high transcriptional variation observed with random integration and the low industrial relevance of transient expression by providing a flexible platform for mAb expression from a consistent clonal background. This work applies a novel SSI-based expression system capable of generating isogenic cell pools in less than 1 month to systematically compare the expression of ten sequence variants of two therapeutically relevant mAbs from two genomic loci under industrially relevant culture conditions. Eight single amino acid mutations in trastuzumab resulted in reduced productivity compared to the wild-type mAb in batch cultures, and three mutations maintained a low-expressing phenotype in fed-batch cultures. The mutations resulted in variant-specific patterns of decreased domain stability and increased ER stress. The application of industrially relevant SSI systems in developability workflows could strengthen the understanding of the sequence determinants of mAb expression to improve mAb design, candidate selection, and process development decisions.

单克隆抗体(mAb)一级序列的变异会对其在生物制药制造平台中的行为产生负面影响,并且鉴定具有较差“可开发性”特征的mAb的努力缺乏从工业相关平台评估mAb表达的可靠方法。中国仓鼠卵巢(CHO)细胞基于位点特异性整合(SSI)平台的最新进展,可以通过提供一个灵活的平台,从一致的克隆背景中表达mAb,从而缓解随机整合中观察到的高转录变异和瞬时表达的低工业相关性。这项工作应用了一种新的基于ssi的表达系统,该系统能够在不到1个月的时间内产生等基因细胞池,以系统地比较来自两个基因组位点的两个治疗相关单克隆抗体在工业相关培养条件下的十个序列变体的表达。与野生型单抗相比,曲妥珠单抗中的8个单氨基酸突变导致分批培养中的产量降低,并且3个突变在饲料分批培养中保持低表达表型。突变导致结构域稳定性下降和内质网胁迫增加的变异特异性模式。工业相关SSI系统在可开发性工作流程中的应用可以加强对单抗表达序列决定因素的理解,从而改进单抗设计、候选物选择和工艺开发决策。
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引用次数: 0
Tailoring Plasmid Design Based on Chain Expression in Cell Line Development for Enhanced Monoclonal and Bispecific Antibody Production 基于链表达的细胞系定制质粒设计增强单克隆和双特异性抗体的产生
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-13 DOI: 10.1002/biot.70104
Shenghai Liu, Qianqian Chen, Lujia Peng, Hongli Li, Bin Zhao, Mengxiao Wu, Yanshen Kang, Ting Hu, Xuefeng Guo, Yanjing Cao, April Xu, Kyu-Sung Lee, Zheng Zhang, Jing Song

The development of robust, high-yielding cell lines represents a critical factor in establishing efficient and cost-effective manufacturing processes within the competitive biopharmaceutical industry. While extensive research has focused on optimizing plasmid design, host cell characteristics, and selection strategies to enhance cell line development, this study specifically investigates strategic expression plasmid design to improve both productivity and product quality of therapeutic proteins in Chinese Hamster Ovary (CHO) cells using transposon technology. Our findings demonstrate that increasing light chain and heavy chain copy numbers while maintaining balanced expression significantly enhances productivity and mitigates purity risks. Furthermore, we reveal that consolidating all chains of asymmetric molecules into a single plasmid, rather than distributing them across multiple plasmids, substantially improves both productivity and purity. For molecules exhibiting poor purity due to imbalanced chain expression, we demonstrate that incorporating an additional copy of the under-expressed chain can yield significant purity improvement. Based on these findings, we propose a customized plasmid design and pool development workflow to ensure high success rates for cell lines producing structurally complex molecules.

在竞争激烈的生物制药行业中,开发健壮、高产的细胞系是建立高效和具有成本效益的制造工艺的关键因素。虽然大量的研究集中在优化质粒设计、宿主细胞特性和选择策略以促进细胞系发育,但本研究专门研究了策略性表达质粒设计,以利用转座子技术提高中国仓鼠卵巢(CHO)细胞治疗性蛋白的产量和产品质量。我们的研究结果表明,在保持平衡表达的同时增加轻链和重链拷贝数可以显著提高生产率并降低纯度风险。此外,我们发现将所有不对称分子链整合到单个质粒中,而不是将它们分布在多个质粒中,大大提高了生产率和纯度。对于由于链表达不平衡而纯度较低的分子,我们证明了添加低表达链的额外拷贝可以显著提高纯度。基于这些发现,我们提出了一种定制的质粒设计和池开发工作流程,以确保细胞系生产结构复杂分子的高成功率。
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引用次数: 0
A Paper-Based Human Kidney Proximal Tubule-on-a-Chip for Efficacy of SGLT2 Inhibitors and Methotrexate-Induced Nephrotoxicity Assessment 基于纸张的人肾近端芯片小管SGLT2抑制剂的疗效和甲氨蝶呤诱导的肾毒性评估
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-11 DOI: 10.1002/biot.70099
Hui Liu, Gui-Mu Guo, Zi-Wei Yu, Yi-Lan Lin, Cui-Hong Lin, Meng-Meng Liu

The human kidney proximal tubule is responsible for glucose reabsorption and serves as a primary target for exogenous toxins. While conventional in vitro cell-based models offer cost-effective alternatives to animal testing, they often fail to replicate the structural and functional complexity of the native proximal tubule. Here, we developed a paper-based human kidney proximal tubule-on-a-chip that mimicked key physiological functions, bridging between traditional cell cultures and animal models. Utilizing porous paper, the chip recreated an in vivo–like three-dimensional microenvironment that supported proximal tubule-specific functions and reproduced essential physiological processes including dynamic glycogen metabolism, glucose reabsorption, and drug transport. The model enabled precise pharmacodynamics evaluation of sodium-glucose co-transporter 2 (SGLT2) inhibitors, yielding median effect concentrations of 0.954 ng/mL for dapagliflozin and 2.685 ng/mL for canagliflozin. The platform maintained consistently high glucose reabsorption inhibition rates (94.59%–95.03%) under different conditions following SGLT2 inhibitors treatment. Furthermore, the methotrexate (MTX)–induced nephrotoxicity evaluation was performed by MTT assay, LDH assay, and glucose reabsorption measurements. The chip accurately reproduced MTX transport dynamics, demonstrating its potential for pharmacokinetic studies. Thus, the paper-based model serves as a reliable platform for pharmacokinetic and nephrotoxicity assessments, offering a valuable tool to replace animal testing and support Reduce, Refine, and Replace experimentation.

人体肾近端小管负责葡萄糖的再吸收,是外源性毒素的主要靶点。虽然传统的体外细胞模型为动物实验提供了经济有效的替代方案,但它们往往无法复制天然近端小管的结构和功能复杂性。在这里,我们开发了一种基于纸张的人类肾脏近端小管芯片,模拟了关键的生理功能,在传统细胞培养和动物模型之间架起了桥梁。利用多孔纸,该芯片重建了一个类似于体内的三维微环境,支持近端小管特异性功能,并重现了包括动态糖原代谢、葡萄糖重吸收和药物运输在内的基本生理过程。该模型能够精确评估钠-葡萄糖共转运蛋白2 (SGLT2)抑制剂的药效学,得出达格列净的中位效应浓度为0.954 ng/mL,卡格列净为2.685 ng/mL。在SGLT2抑制剂治疗的不同条件下,该平台始终保持较高的葡萄糖重吸收抑制率(94.59%-95.03%)。此外,通过MTT法、LDH法和葡萄糖重吸收法评估甲氨蝶呤(MTX)诱导的肾毒性。该芯片精确地再现了MTX转运动力学,证明了其在药代动力学研究中的潜力。因此,基于纸张的模型可作为药代动力学和肾毒性评估的可靠平台,为替代动物试验提供了有价值的工具,并支持减少,改进和替代实验。
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引用次数: 0
3D Printed Microfluidic Chromatographic Column for Fast Downstream Processing Development 用于快速下游加工开发的3D打印微流控色谱柱
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-10 DOI: 10.1002/biot.70095
Vladimir Matining, Mario Messina, Benedetta Sechi, Davide Moscatelli, Mattia Sponchioni

3D printing is emerging as a promising fabrication technique for microfluidic devices. In this work, this technology was exploited in the development of a microfluidic chromatographic column with nominal volume of 54 µL. The microcolumn was packed with a cation exchange resin and characterized, using potassium iodide as a tracer, in terms of porosity (ε = 0.72), plate number, and asymmetry factor (0.8 < AS < 1.8 for flowrates >50 µL/min). To showcase the potential of this microdevice, it was exploited in the characterization of the chromatographic behavior of lysozyme. The measured saturation capacity (q= 88.14 g/Lresin at 340 cm/h) was in line with the manufacturer declaration (85–135 g/L at <500 cm/h). In addition, the effect of NaCl at different concentrations on the protein adsorption isotherm was characterized, demonstrating a Langmuir to anti-Langmuir transition at concentrations ≥300 mM. The axial dispersion coefficient was finally determined (DAX${{mathcal{D}}_{AX}}$= 6.7 · 10−9 m2/s). In this way, the mcirofluidic column allowed to develop a comprehensive mechanistic model describing the transport of lysozyme in the chromatographic medium using only 30 µL of resin and <1 g of protein, addressing the issue of limited availability of biomolecules and streamlining the process development.

3D打印正在成为一种很有前途的微流体器件制造技术。在这项工作中,该技术被用于开发一种标准体积为54 μ L的微流控色谱柱。用阳离子交换树脂填充微柱,用碘化钾作为示踪剂,对微柱的孔隙率(ε = 0.72)、板数和不对称系数(0.8 <;& lt;1.8流速>;50 μ L/min)。为了展示这种微型装置的潜力,它被用于表征溶菌酶的色谱行为。测量的饱和容量(q∞= 88.14 g/L树脂,340 cm/h)符合制造商声明(85-135 g/L, <500 cm/h)。此外,研究了不同浓度NaCl对蛋白质吸附等温线的影响,结果表明,NaCl浓度≥300 mM时存在Langmuir向反Langmuir转变。最后确定了轴向分散系数(D a X ${{mathcal{D}}_{AX}}$ = 6.7·10−9 m2/s)。通过这种方式,微流控柱可以建立一个全面的机制模型,描述溶菌酶在色谱介质中的运输,仅使用30µL树脂和1g蛋白质,解决了生物分子可用性有限的问题,并简化了工艺开发。
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引用次数: 0
AI-Driven Quality Monitoring and Control in Stem Cell Cultures: A Comprehensive Review 人工智能驱动的干细胞培养质量监测和控制:综合综述
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-10 DOI: 10.1002/biot.70100
Rohan Singh, Hamid Ebrahimi Orimi, Praveen Kumar Raju Pedabaliyarasimhuni, Corinne A. Hoesli, Moncef Chioua

Recent advancements in stem cell research forge them into one of the most promising sources for cell therapy applications. Quality monitoring in stem cell culture is essential for ensuring consistency, viability, and therapeutic efficacy. Traditional methods involve periodic sampling for conducting endpoint assays such as cell viability, proliferation, and differentiation using microscopy and flow cytometry, which are labor-intensive and often lack the real-time monitoring of the processes for scale-up applications. This paper explores artificial intelligence (AI)-driven approaches for real-time quality control, integrating machine vision, predictive modeling, and sensor-based monitoring. AI models analyze high-resolution imaging and multi-sensor data to dynamically track critical quality attributes (CQAs), including cell morphology, proliferation rate, differentiation potential, environmental stability (pH, oxygen, and nutrient levels), genetic integrity, and contamination risks. These models enable automated anomaly detection, differentiation tracking, and adaptive culture optimization. By leveraging real-time feedback systems and multi-omics integration, AI-driven techniques enhance scalability, reproducibility, and process automation in stem cell biomanufacturing. This review outlines current advancements, challenges, and future directions in AI-assisted quality monitoring and highlights its potential to improve fully automated, scalable production of stem cell lines for clinical translation and regulatory compliance in regenerative medicine.

干细胞研究的最新进展使其成为细胞治疗应用中最有前途的来源之一。干细胞培养的质量监测对于确保一致性、活力和治疗效果至关重要。传统的方法包括使用显微镜和流式细胞术定期取样进行终点分析,如细胞活力、增殖和分化,这是劳动密集型的,并且通常缺乏对大规模应用过程的实时监控。本文探讨了人工智能(AI)驱动的实时质量控制方法,集成了机器视觉、预测建模和基于传感器的监测。人工智能模型分析高分辨率成像和多传感器数据,以动态跟踪关键质量属性(cqa),包括细胞形态、增殖率、分化潜力、环境稳定性(pH值、氧气和营养水平)、遗传完整性和污染风险。这些模型支持自动异常检测、差异跟踪和自适应培养优化。通过利用实时反馈系统和多组学集成,人工智能驱动的技术增强了干细胞生物制造的可扩展性、可重复性和过程自动化。本文概述了人工智能辅助质量监测的当前进展、挑战和未来方向,并强调了人工智能在提高临床转化和再生医学法规遵从性的全自动化、可扩展的干细胞系生产方面的潜力。
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引用次数: 0
Chromosomal Integration of budAB Operons and Pathway Rewiring Enhance Acetoin Production From Starch in Vibrio diabolicus budAB操纵子的染色体整合和途径重布线促进了双歧弧菌淀粉乙酰素的产生
IF 3.1 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-08-10 DOI: 10.1002/biot.70094
Yuan He, Guoli Lian, Ning Guo, Zheng-Jun Li

Acetoin is a key platform chemical with diverse industrial applications. In this study, the marine bacterium Vibrio diabolicus, characterized by its rapid growth and strong ability to utilize starch, was systematically engineered for efficient conversion of starch into acetoin. A suicide plasmid-mediated homologous recombination system was first developed to investigate the roles of four endogenous amylase genes. Based on transcriptomic analysis, two strong constitutively active endogenous promoters were identified and functionally validated to enhance gene expression. To increase acetoin production, the 2,3-butanediol dehydrogenase gene and polyhydroxyalkanoate synthase gene were deleted, thereby eliminating carbon flux into competing pathways for 2,3-butanediol and poly-3-hydroxybutyrate biosynthesis. Subsequently, multiple copies of the budAB operon were integrated into the chromosome to strengthen the acetoin biosynthetic route. The final engineered strain produced 13.21 g/L of acetoin within 12 h of shake flask cultivation, reflecting a significant enhancement in production efficiency. This study presents the first successful case of metabolic engineering in V. diabolicus for direct and efficient production of acetoin from starch, highlighting its significant potential for industrial-scale bioproduction.

乙托因是一种具有多种工业应用的关键平台化学品。本研究对生长速度快、对淀粉利用能力强的海洋细菌diabolicus弧菌进行了系统工程改造,使其能将淀粉高效转化为乙酰胆碱。首先建立了一种自杀质粒介导的同源重组系统来研究四种内源性淀粉酶基因的作用。基于转录组学分析,鉴定了两个强组成活性内源性启动子,并对其进行了功能验证,以增强基因表达。为了提高乙酰丙酮的产量,2,3-丁二醇脱氢酶基因和聚羟基烷酸酯合成酶基因被删除,从而消除了碳通量进入2,3-丁二醇和聚3-羟基丁酸酯生物合成的竞争途径。随后,budAB操纵子的多个拷贝被整合到染色体中,以加强乙酰蛋白的生物合成途径。在摇瓶培养12 h内,最终工程菌株的乙酰素产量为13.21 g/L,生产效率显著提高。本研究首次成功地利用代谢工程技术从淀粉中直接高效地生产乙酰胆碱,突出了其在工业规模生物生产方面的巨大潜力。
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