Biosensors and Bioelectronics最新文献_第6页

Microfluidic device for continuous blood plasma separation from whole blood 全血血浆连续分离的微流控装置。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-17 DOI: 10.1016/j.bios.2026.118404

Surendran Velmurugan , Melis Güler Girbas , Can Dincer , Meltem Avci-Adali , Stefan Partel

Sample preparation is a critical step in clinical diagnostics, as plasma must be separated from whole blood before analysis. Conventional centrifugation is time-consuming, require bulky equipment, and is unsuitable for point-of-care (POC) applications. Here, we present a simple and low-cost microfluidic device that achieves continuous (uninterrupted flow separation without batch processing) and real-time (immediate flow separation and inline without delays) plasma separation from undiluted whole human blood without external forces. Six channel geometries were evaluated by numerical simulations to investigate the effects of side-channel orientation and channel height on red blood cell (RBC) contamination carryover. The optimized configuration with a 20-μm channel height, minimized recirculation zones and enhanced inertial focusing, leading to reduced RBC contamination in the side channel plasma outlet. Devices were then fabricated by standard photolithography and soft lithography. Experimental validation with whole human blood demonstrated that the device significantly reduced red blood cell (RBC) contamination across a flow rate range of 0.05–0.2 mL min⁻¹, yielding plasma with substantially lower RBC levels compared with whole blood, although low residual RBCs were still present. Flow cytometry further confirmed an efficient depletion of peripheral blood mononuclear cells (PBMCs), resulting in markedly lower leukocyte content than in plasma obtained by standard centrifugation. The plasma yield reached 3.8 %, corresponding to an extraction rate of 6.5 μL min⁻¹, and hemolysis levels remained comparable to centrifuged samples, indicating preservation of plasma integrity. This established device provides a robust platform for high-purity plasma separation and has a strong potential for point-of-care diagnostic applications.

样品制备是临床诊断的关键步骤，因为血浆必须在分析前从全血中分离出来。传统的离心费时，需要笨重的设备，并且不适合护理点（POC）应用。在这里，我们提出了一种简单和低成本的微流体装置，可以实现连续（不间断流动分离，无需批量处理）和实时（即时流动分离，在线无延迟）的血浆分离，无需外力。通过数值模拟对六种通道几何形状进行了评估，以研究通道侧方向和通道高度对红细胞污染携带的影响。优化后的通道高度为20 μm，减少了再循环区域，增强了惯性聚焦，从而减少了侧通道等离子体出口的RBC污染。然后用标准光刻法和软光刻法制造器件。用全血进行的实验验证表明，该装置在0.05-0.2 mL min-1的流速范围内显著减少了红细胞（RBC）污染，与全血相比，产生的血浆中红细胞水平显著降低，尽管仍存在低残留红细胞。流式细胞术进一步证实了外周血单个核细胞（PBMCs）的有效耗竭，导致白细胞含量明显低于标准离心获得的血浆。血浆得率达到3.8%，相当于6.5 μL min-1的提取率，溶血水平与离心后的样品相当，表明血浆的完整性得到了保存。这种已建立的设备为高纯度血浆分离提供了一个强大的平台，并且在即时诊断应用方面具有强大的潜力。

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引用次数: 0

Biomimetic self-sustained ion oscillation in miscibility gap and chemical inductance impact in potassium sensing 混溶间隙的仿生自维持离子振荡及化学电感对钾传感的影响。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-17 DOI: 10.1016/j.bios.2026.118413

Marini L , Roop L. Mahajan , Chithra Lekha P , T. Devasena

A biomimetic potassium ion-selective electrode (ISE) was developed using a graphene oxide (GO)-coated nickel foam (NF) substrate functionalized with a polyethylene glycol (PEG)-based ion-selective layer (ISL). The sensing mechanism centers on a miscibility gap formed within the ISL due to PEG's amphiphilic nature when complexed with potassium iodide—driving phase separation into hydrophilic and hydrophobic domains. This phase heterogeneity introduces interfacial barriers that delay ionic relaxation and facilitate self-sustained ion oscillation, as evidenced by low-frequency inductive behavior in electrochemical impedance spectroscopy (EIS)—demonstrating a chemical inductor effect. The design eliminates the need for costly ionophores, such as valinomycin, presenting a scalable and cost-effective sensing platform. The sensor exhibited a linear response to K⁺ concentrations ranging from 0.5 to 10 mM, with a high sensitivity of 14.09 × 10² mA/mM·cm² and a detection limit of 0.69 mM. Selectivity tests showed minimal interference from Na⁺, Ca²⁺, urea, NH₄⁺, and ascorbic acid ions, and the sensor demonstrated excellent stability over 100 cycles. These findings provide a robust framework for developing low-cost, biomimetic electrochemical potassium sensing.

采用聚乙二醇（PEG）基离子选择层（ISL）功能化的氧化石墨烯（GO）涂层泡沫镍（NF）衬底，开发了一种仿生钾离子选择电极（ISE）。当与碘化钾驱动相分离成亲水和疏水结构域时，由于PEG的两亲性，在ISL内形成了一个混溶间隙，这是传感机制的核心。正如电化学阻抗谱（EIS）中的低频感应行为所证明的那样，这种相非均质性引入了界面障碍，延迟了离子弛豫并促进了离子的自我持续振荡——证明了化学电感效应。该设计消除了对昂贵的离子载体（如valinomycin）的需求，提供了一个可扩展且具有成本效益的传感平台。灵敏度为14.09 × 102 mA/mM·cm2，检出限为0.69 mM。选择性测试表明，Na+、Ca2+、尿素、NH4+和抗坏血酸离子对传感器的干扰最小，且传感器在100次循环内具有良好的稳定性。这些发现为开发低成本、仿生电化学钾传感提供了一个强有力的框架。

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引用次数: 0

LDOB: multi-dimensional programmable lactose-derived oligosaccharide biosensors LDOB：多维可编程乳糖衍生低聚糖生物传感器。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-16 DOI: 10.1016/j.bios.2026.118407

Li-Hua Liu , Bo Xu , Ying Huang , Rongrong Chen , Yunxia Li , Yu Zhang , Hong Huang , Kuo Zhang , Junyang Huang , Ao Jiang

Human milk oligosaccharides (HMOs) are the third most abundant solid constituent of breast milk and exert anti-infective, prebiotic and immunomodulatory functions. Microbial de novo synthesis is currently the only scalable route for commercial HMOs production, yet the absence of rapid and universal analytical tools has become a major bottleneck for strain improvement. Exploiting the strict stoichiometry between lactose consumption and HMOs biosynthesis, we constructed a set of Escherichia coli-based lactose-derived oligosaccharide biosensors (LDOB)—genetically engineered whole-cell systems that translate lactose concentration into inversely correlated biomass and fluorescence read-outs, enabling real-time screening of HMO-producing strains without downstream metabolite processing. The sensing circuit integrates multidimensional negative-feedback modules—encompassing multi-repression, targeted protein degradation and an occluded ribosome binding site (oRBS)—endowing LDOB with a wide dynamic range, straightforward operability, excellent compatibility, and substantial application potential. Validation experiments demonstrated that LDOB signals exhibited strong consistency with the yields of key HMOs, including 2′-fucosyllactose (2′-FL) and lacto-N-neotetraose (LNnT). Furthermore, coupling LDOB with fluorescence-activated droplet sorting (FADS) enabled high-throughput screening of evolved strains, resulting in remarkable titer enhancements of 42.8 % for 2′-FL and 86.4 % for LNnT, respectively. Collectively, these findings fully confirm the superiority of LDOB in monitoring HMOs synthesis and screening high-yield HMOs-producing strains, providing a valuable tool for advancing HMOs biomanufacturing.

人乳寡糖（HMOs）是母乳中含量第三高的固体成分，具有抗感染、益生元和免疫调节功能。微生物从头合成是目前商业HMOs生产中唯一可扩展的途径，但缺乏快速和通用的分析工具已成为菌株改进的主要瓶颈。利用乳糖消耗和HMOs生物合成之间的严格化学计量学，我们构建了一套基于大肠杆菌的乳糖衍生寡糖生物传感器(LDOB)-基因工程全细胞系统，将乳糖浓度转化为负相关的生物量和荧光读数，从而能够实时筛选产生hmo的菌株，而无需下游代谢物处理。传感电路集成了多维负反馈模块，包括多重抑制、靶向蛋白质降解和封闭核糖体结合位点（oRBS），使LDOB具有广泛的动态范围、简单的可操作性、出色的兼容性和巨大的应用潜力。验证实验表明，LDOB信号与关键HMOs的产率具有很强的一致性，包括2'- focusyllactose （2'- fl）和lactto -n -neotetraose （LNnT）。此外，将LDOB与荧光激活液滴分选（FADS）相结合，实现了进化菌株的高通量筛选，2'-FL和LNnT的滴度分别提高了42.8%和86.4%。综上所述，这些发现充分证实了LDOB在监测HMOs合成和筛选高产HMOs生产菌株方面的优势，为推进HMOs生物制造提供了有价值的工具。

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引用次数: 0

Bulge DNA-driven CRISPR/Cas12a dynamic activation circuit enables highly sensitive and versatile biosensing 凸起dna驱动的CRISPR/Cas12a动态激活电路实现高灵敏度和多用途的生物传感

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-16 DOI: 10.1016/j.bios.2026.118412

Luyu Wei , Ziqi Cheng , Mingrui Xu , Hengye Chen , Wei Lan , Wanjun Long , Yuanbin She , Haiyan Fu

CRISPR/Cas12a has emerged as an innovative biosensing tool; however, its intrinsically linear accumulation of cleavage signals limits detection sensitivity. Herein, we report a bulge DNA (BD)-driven CRISPR/Cas12a dynamic activation circuit, termed CBD, as a highly sensitive and versatile biosensing platform for both nucleic acid and non-nucleic acid targets. The BD structure was rationally engineered to undergo programmable structural and functional transformation upon bulge degradation, thereby initiating exponential, self-amplifying activation of the Cas12a circuit. For nucleic acid targets, direct Cas12a recognition triggers BD cleavage and a positive feedback loop, enabling highly sensitive detection with a limit of 14 CFU/mL for methicillin-resistant Staphylococcus aureus. For non-nucleic acid targets, a universal single-stranded DNA activator was linked to the aptamer-complementary strand, enabling target-responsive release and subsequent initiation of the CBD system without altering the crRNA or BD sequence. This strategy enabled the detection of pesticides and mycotoxins at the picogram-per-milliliter level. Furthermore, an “OR” logic gate was constructed for the simultaneous detection of dual mycotoxins, highlighting the platform's capability for multiplexed hazard monitoring. Overall, CBD demonstrates significant potential as a new paradigm for next-generation biosensing technologies.

CRISPR/Cas12a已经成为一种创新的生物传感工具；然而，其解理信号固有的线性积累限制了检测灵敏度。在此，我们报告了一个凸起DNA （BD）驱动的CRISPR/Cas12a动态激活电路，称为CBD，作为核酸和非核酸靶点的高度敏感和通用的生物传感平台。合理设计BD结构，使其在鼓包降解时进行可编程的结构和功能转换，从而启动Cas12a电路的指数级自放大激活。对于核酸靶标，直接识别Cas12a触发BD切割和正反馈循环，对耐甲氧西林金黄色葡萄球菌的检测灵敏度高，限限为14 CFU/mL。对于非核酸靶标，将通用单链DNA激活子连接到适体互补链上，在不改变crRNA或BD序列的情况下，实现靶标响应释放和随后启动CBD系统。这一策略使检测到的农药和真菌毒素在每毫升微克的水平。此外，构建了一个“或”逻辑门，用于同时检测双真菌毒素，突出了该平台的多路危害监测能力。总体而言，CBD作为下一代生物传感技术的新范式显示出巨大的潜力。

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引用次数: 0

A novel ratiometric biosensor based on germanene nanosheets for rapid and sensitive BRCA1 detection in differentiating T cells 一种基于锗烯纳米片的新型比例生物传感器，用于快速灵敏地检测分化T细胞中的BRCA1。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-16 DOI: 10.1016/j.bios.2026.118411

Yingying Huang , Shu Xiao , Zhiming Zhong , Kamning Yuen , Bohan Yin , Jiareng Chen , Jiaxiang Yan , Jun Peng , Qin Zhang , Fengqi Wang , Bin Liu , Siu Hong Dexter Wong , Mo Yang

The BRCA1 gene is essential for the developmental regulation and function of T lymphocytes, yet its mRNA expression during T-cell differentiation remains unclear due to the lack of effective in-situ monitoring tools. To address this, we developed a ratiometric fluorescent nanosensor based on Förster resonance energy transfer (FRET) for reliable quantification of BRCA1 mRNA in living CD8⁺ T cells. The sensor comprises a fluorescein-labeled DNA probe (FAM_cDNA) assembled with Triton X-100-modified methylgermanene nanosheets (GeT), forming an efficient FRET pair. In the absence of the target, FAM_cDNA adsorbs onto the GeT surface, resulting in a FRET effect that quenches FAM fluorescence and enhances GeT emission by 1.63-fold. Upon specific hybridization with BRCA1 mRNA, the probe detaches, disrupting the FRET process and causing a quantitative ratiometric shift (I₅₂₀/I₆₄₀). This self-calibrating system demonstrates high sensitivity, with detection limits of 18.1 pM (R² = 0.985) for synthetic DNA and 17.2 pM (R² = 0.996) for mRNA, and a rapid response time (∼10 min). Importantly, the nanoprobe enabled ratiometric imaging of endogenous BRCA1 mRNA in living CD8⁺ T cells, revealing a significant increase in the I₅₂₀/I₆₄₀ ratio during activation, visually confirming BRCA1 upregulation consistent with RNA-seq data. This work provides a robust assay for T-cell studies and highlights red-emissive germanene as a promising platform for ratiometric biosensing.

BRCA1基因对T淋巴细胞的发育调控和功能至关重要，但由于缺乏有效的原位监测工具，其mRNA在T细胞分化过程中的表达尚不清楚。为了解决这个问题，我们开发了一种基于Förster共振能量转移（FRET）的比例荧光纳米传感器，用于可靠地定量活CD8+ T细胞中的BRCA1 mRNA。该传感器由荧光素标记的DNA探针（FAMcDNA）与Triton x -100修饰的甲基锗烯纳米片（GeT）组装而成，形成一个高效的FRET对。在没有目标物的情况下，FAMcDNA吸附在GeT表面，产生FRET效应，猝灭FAM荧光，使GeT发射增强1.63倍。在与BRCA1 mRNA特异性杂交后，探针分离，破坏FRET过程并导致定量比率偏移（I520/I640）。该自校准系统具有很高的灵敏度，对合成DNA的检出限为18.1 pM (R2 = 0.985)，对mRNA的检出限为17.2 pM (R2 = 0.996)，并且反应时间快速（约10分钟）。重要的是，纳米探针能够在活的CD8+ T细胞中实现内源性BRCA1 mRNA的比例成像，揭示了I520/I640比值在激活过程中显著增加，从视觉上证实了BRCA1上调与RNA-seq数据一致。这项工作为t细胞研究提供了一个强大的分析方法，并强调了红发射锗烯作为一个有前途的比率生物传感平台。

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引用次数: 0

Synergistic coupling of a spherical nucleic acid DNA walker and a redox cascade for self-calibrated ratiometric detection of endocrine disruptors 球形核酸DNA助行器和氧化还原级联的协同耦合用于自校准比例检测内分泌干扰物

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-16 DOI: 10.1016/j.bios.2026.118372

Na Wu , Yijie Chen , Lu Shi , Weiheng Liu , Jianping Xu , Chenqi Niu , Huaping Peng

Endocrine-disrupting chemicals (EDCs), such as 17β-estradiol (E2) and bisphenol A (BPA), are pervasive environmental contaminants that pose significant health risks even at trace concentrations. Their on-site monitoring remains challenging due to insufficient sensitivity and stability of conventional detection methods. To address this, we developed a universal ratiometric fluorescence biosensor by integrating a target-activated spherical nucleic acid (SNA) with an enzyme-nanozyme- mediated redox cascade for the ultrasensitive and self-calibrating detection of EDCs. The sensing mechanism is initiated by aptamer recognition, which triggers a Nb. BbvCI-powered DNA walking circuit on gold nanoparticles (AuNPs), resulting in the release of numerous alkaline phosphatase (ALP) molecules. These ALP then catalyze the conversion of ascorbic acid 2-phosphate (AAP) to ascorbic acid (AA), which subsequently reduces CoOOH nanoflakes (CoOOH NFs) in a g-C₃N₄/CoOOH nanohybrid. This redox cascade quenches the oxidase-mimicking activity of CoOOH NFs, suppresses oxidation-mediated emission at 556 nm, and simultaneously restores the intrinsic fluorescence of g-C₃N₄ nanosheets at 448 nm. This anti-correlated dual-signal output enables accurate and stable quantification against environmental interference. The biosensor achieved detection limits of 100 pM for E2 and 240 pM for BPA, outperforming conventional colorimetric methods by 2 orders of magnitude. Furthermore, the practical applicability was successfully validated using real water samples yielding high recovery rates and demonstrating excellent reliability. By synergizing SNA amplification with an enzyme-nanozyme-modulated ratiometric sensing strategy, this work provides a robust and generalizable platform for the on-site screening of trace-level EDCs, holding great promise for applications in environmental monitoring and public health diagnostics.

内分泌干扰化学物质（EDCs），如17β-雌二醇（E2）和双酚A (BPA)，是普遍存在的环境污染物，即使是微量浓度也会对健康造成重大威胁。由于传统检测方法的灵敏度和稳定性不足，它们的现场监测仍然具有挑战性。为了解决这个问题，我们开发了一种通用的比率荧光生物传感器，通过将靶激活的球形核酸（SNA）与酶-纳米酶介导的氧化还原级联相结合，用于超灵敏和自校准的EDCs检测。感应机制由适体识别启动，从而触发Nb。bbvci驱动的DNA行走电路在金纳米颗粒（AuNPs）上，导致大量碱性磷酸酶（ALP）分子的释放。这些ALP随后催化抗坏血酸2-磷酸（AAP）转化为抗坏血酸（AA），随后在g-C3N4/CoOOH纳米杂化物中还原CoOOH纳米片（CoOOH NFs）。该氧化还原级联抑制了CoOOH NFs的氧化酶模拟活性，抑制了556 nm的氧化介导发射，同时恢复了g-C3N4纳米片在448 nm的固有荧光。这种抗相关双信号输出使精确和稳定的量化对抗环境干扰。该生物传感器对E2的检测限为100 pM，对BPA的检测限为240 pM，比传统比色法高出2个数量级。此外，使用实际水样成功验证了实际适用性，获得了高回收率，并展示了出色的可靠性。通过SNA扩增与酶-纳米酶调节的比例传感策略的协同作用，本研究为痕量EDCs的现场筛选提供了一个强大且可推广的平台，在环境监测和公共卫生诊断中具有很大的应用前景。

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引用次数: 0

Multiple microRNAs analysis based on DNAzyme cascade DNA fiber barcodes for cell typing 基于DNAzyme级联DNA纤维条形码的多microRNAs分析用于细胞分型。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-15 DOI: 10.1016/j.bios.2026.118383

Xuewei Li , Ji Zheng , Xiaoxing Fang , Wenqi Gao , Hejiang Chen , Zhenyi Liu , Chao Li , Ruiping Zhang

Accurate tumor subtyping through multiplex miRNA profiling is critical for precision medicine but remains challenging due to limitations in current methods, including cross-reactivity, cost, and stability. Herein, we present a DNAzyme-powered DNA fiber barcode platform that combines G-quadruplex (G4)/hemin catalysis, miRNA-responsive displacement, and polydopamine (PDA)-mediated fluorescence modulation for sensitive and specific miRNA detection. The system operates via a target-dependent “switch”: in the absence of target miRNAs, G4/hemin DNAzymes catalyze dopamine (DA) oxidation into PDA that efficiently quench fluorescence through Förster resonance energy transfer (FRET); when target miRNAs are present, they competitively binding to the capture strands on DNA fibers with higher affinity, displacing the G4 structures, inhibiting DNAzyme formation and preserving the fluorescence signal, with fluorescence intensity showing a positive correlation with target concentration. This mechanism enables dual qualitative and quantitative analysis with a broad linear range (50 pM-100 nM) and low detection limits (5.50 pM). Key advantages include stable, tunable G4/hemin transduction, enhanced kinetics and signal-to-noise through synergistic displacement-quenching, and multicolor barcoding for one-pot multiplex miRNA detection. Validated against qPCR with high concordance, this platform overcomes existing technical barriers to enable robust miRNA-based cell typing classification and precision diagnostics.

通过多重miRNA分析准确的肿瘤亚型对精准医疗至关重要，但由于现有方法的局限性，包括交叉反应性、成本和稳定性，仍然具有挑战性。在此，我们提出了一个dnazyme驱动的DNA纤维条形码平台，该平台结合了g -四重体(G4)/血红素催化、miRNA响应位移和多多巴胺（PDA）介导的荧光调制，用于敏感和特异性的miRNA检测。该系统通过目标依赖的“开关”运行：在缺乏目标mirna的情况下，G4/hemin DNAzymes催化多巴胺（DA）氧化成PDA，通过Förster共振能量转移（FRET）有效地猝灭荧光；当目标mirna存在时，它们以更高的亲和力与DNA纤维上的捕获链竞争性结合，取代G4结构，抑制DNAzyme的形成并保留荧光信号，荧光强度与靶浓度呈正相关。该机制可实现双定性和定量分析，具有宽线性范围（50 pM-100 nM）和低检出限（5.50 pM）。主要优势包括稳定，可调的G4/hemin转导，通过协同位移猝灭增强动力学和信号到噪声，以及用于一锅多重miRNA检测的多色条形码。通过高一致性的qPCR验证，该平台克服了现有的技术障碍，实现了基于mirna的细胞分型分类和精确诊断。

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引用次数: 0

A dual-emission near-infrared fluorogenic probe for sensing Catechol-O- methyltransferase activity from in vitro to in vivo 一种双发射近红外荧光探针，用于检测从体外到体内的儿茶酚- o -甲基转移酶活性。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-15 DOI: 10.1016/j.bios.2026.118409

Fang-Yuan Wang , Lai Wei , Pu Wang , Wen-Chao Chen , Zu-Jia Chen , Zi-Qiong Zhou , Guang-Hao Zhu , Qing Gong , Lu-Ping Qin (∗∗∗∗) , Ling Yang (∗∗∗) , Guang-Bo Ge (∗∗) , Ping Wang (∗)

Human catechol-O-methyltransferase (COMT, EC 2.1.1.6) plays a key role in neurotransmitter signaling and metabolism in the central nervous system. Optical sensing of COMT activity provides a powerful approach to study its biological functions, however, existing fluorescent probes are limited by short emission wavelengths and low sensitivity. Herein, we present TCFC, the first dual-emission near-infrared (NIR) fluorogenic substrate for COMT, rationally designed via a structure-modulated strategy. Under physiological conditions, TCFC was rapidly O-methylated by COMT to yield product 8-MTCFC, which exhibited strong fluorescence at 620 nm and 695 nm upon excitation at 470 nm and 610 nm, respectively. The dual-emission property of TCFC enhanced detection sensitivity and versatility, enabling accurate assessment of COMT activity across individuals and species while supporting high-throughput screening of potential COMT inhibitors. Moreover, the NIR emission of TCFC allowed in situ visualization of COMT activity in living cells and various brain regions of rats with high spatiotemporal resolution, and revealed a reduction of COMT activity in the hippocampal region of a mouse model of Alzheimer's disease, offering insights into its role in neurodegeneration. Collectively, TCFC proved to be a powerful NIR fluorogenic substrate for monitoring COMT activity in complex biological systems, enabling mechanistic studies and high-throughput screening of potential inhibitors.

人儿茶酚o -甲基转移酶（COMT, EC 2.1.1.6）在中枢神经系统的神经递质信号传导和代谢中起关键作用。COMT活性的光学传感为研究其生物学功能提供了强有力的途径，但现有的荧光探针受发射波长短和灵敏度低的限制。在此，我们提出了TCFC，第一个双发射近红外（NIR）荧光基板的COMT，通过结构调制策略合理设计。在生理条件下，TCFC经COMT快速o -甲基化得到产物8-MTCFC，在470 nm和610 nm激发下分别在620 nm和695 nm处表现出强荧光。TCFC的双发射特性增强了检测灵敏度和多功能性，能够准确评估不同个体和物种的COMT活性，同时支持潜在COMT抑制剂的高通量筛选。此外，TCFC的近红外发射可以以高时空分辨率原位可视化大鼠活细胞和大脑各区域的COMT活性，并揭示了阿尔茨海默病小鼠模型海马区域COMT活性的降低，为其在神经退行性变中的作用提供了见解。总的来说，TCFC被证明是一种强大的近红外荧光底物，可用于监测复杂生物系统中的COMT活性，从而进行机制研究和高通量筛选潜在的抑制剂。

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引用次数: 0

Covalently engineered PQDs-fluorescent protein FRET probes for tumor imaging and diagnosis 共价工程pqds荧光蛋白FRET探针用于肿瘤成像和诊断。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-14 DOI: 10.1016/j.bios.2026.118381

Jingli Bi, Lingyun Gao, Yufei Xu, Zhenhai Tan, Shengwang Zhou

Förster Resonance Energy Transfer (FRET)-based biosensors are powerful tools for monitoring biomolecular interactions and microenvironmental dynamics with high precision. Herein, we constructed two distinct FRET probes by covalently assembling fluorescent proteins (mCherry or sfGFP) onto CsPbX₃ PQDs via enzyme-inhibitor-linked surface modification. The FRET probes respond to glutathione (GSH)-mediated reduction through disulfide bond cleavage, leading to modulations in energy transfer efficiency. Upon anti-HER2-mediated internalization, the PQDs-fluorescent protein probes facilitate real-time visualization of dynamic intracellular GSH fluctuations in living cells. The different GSH levels observed between malignant and normal cells serve as a critical biomarker for cancer detection, highlighting the potential of GSH-responsive nanoprobes for integrated diagnostic and therapeutic applications in oncology.

Förster基于共振能量转移（FRET）的生物传感器是高精度监测生物分子相互作用和微环境动力学的强大工具。在这里，我们通过酶抑制剂连接的表面修饰将荧光蛋白（mCherry或sfGFP）共价组装到CsPbX3 pqd上，构建了两个不同的FRET探针。FRET探针响应谷胱甘肽（GSH）介导的还原通过二硫键裂解，导致能量转移效率的调节。在抗her2介导的内化过程中，pqds荧光蛋白探针可以实时可视化活细胞内动态GSH波动。在恶性细胞和正常细胞之间观察到不同的谷胱甘肽水平，作为癌症检测的关键生物标志物，突出了谷胱甘肽反应性纳米探针在肿瘤学综合诊断和治疗应用中的潜力。

引用次数: 0

Programmable PNA–nanoparticle hybrids as nanoscale recognition architectures for amplification-free nucleic acid recognition 可编程的pna -纳米粒子杂交体作为无扩增核酸识别的纳米级识别架构。

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics

Pub Date : 2026-01-14 DOI: 10.1016/j.bios.2026.118380

Satheesh Natarajan , David Skrodzki , Neela H. Yennawar , John Gershenson , Dipanjan Pan

Rapid, field-deployable nucleic acid diagnostics require robust recognition elements and simple signal transduction. We present a programmable peptide nucleic acid–gold nanoparticle (PNA–AuNP) platform for amplification-free detection of Schistosoma spp. DNA. Thiolated PNAs were conjugated to citrate-stabilized AuNPs, forming a nanointerface that translates sequence-specific hybridization into a plasmonic colorimetric signal. Structural and thermodynamic validation confirmed duplex integrity: X-ray photoelectron spectroscopy revealed N 1s (+0.4 eV) and O 1s carbonyl (−0.2 eV) shifts, while FTIR showed amide I/II changes and new sugar–phosphate bands (921–1045 cm⁻¹) with a phosphate fingerprint at 1237 cm⁻¹. Zeta potential shifted from −35 mV (DNA) to −15 mV (PNA–DNA complex), indicating charge neutralization. Isothermal titration calorimetry demonstrated strong binding (Kd ≈ 20.9 nM, ΔH ≈ 252.4 kcal/mol, ΔS ≈ 880.4 cal/mol·K), nearly twice that of ASO–DNA. Guided by these insights, a tri-probe plasmonic lateral flow assay achieved a detection limit of 0.01 ng. mL⁻¹ within 20 min tenfold more sensitive than ASO-based LFAs and comparable to RT-PCR while maintaining high specificity, reproducibility (CV <10 %), and six-week stability. This work establishes PNAs as high-affinity, enzymatically stable probes, offering a versatile framework for rapid, amplification-free diagnostics in resource-limited settings.

快速，现场可部署的核酸诊断需要强大的识别元素和简单的信号转导。我们提出了一种可编程肽核酸-金纳米粒子（PNA-AuNP）平台，用于无扩增检测血吸虫DNA。硫代化的PNAs与柠檬酸盐稳定的aunp结合，形成纳米界面，将序列特异性杂交转化为等离子体比色信号。结构和热力学验证证实了双相完整性：x射线光电子能谱显示N 1s （+0.4 eV）和O 1s羰基（-0.2 eV）位移，而FTIR显示酰胺I/II变化和新的糖-磷酸谱带（921-1045 cm-1），磷酸盐指纹在1237 cm-1。Zeta电位从-35 mV （DNA）转移到-15 mV （PNA-DNA复合物），表明电荷中和。等温滴定量热法显示出较强的结合（Kd≈20.9 nM， ΔH≈252.4 kcal/mol， ΔS≈880.4 cal/mol·K），几乎是ASO-DNA的两倍。在这些见解的指导下，三探针等离子体横向流动分析达到了0.01 ng的检测限。mL-1在20分钟内的敏感性比基于aso的LFAs高10倍，与RT-PCR相当，同时保持高特异性，重复性(CV

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