ACS SensorsPub Date : 2026-10-09DOI: 10.1021/acssensors.6c02645
Lin Hu, Honghua Hu, Fang Liu, Xingrong Li, Qinghua Wu, Xuhuai Fu, Diangeng Li, Xuan Wu, Rong Xiang, Rongzhong Huang, Xiaosong Li, Yi Li
{"title":"Branched Split Activator-Regulated Cas12a Biosensor For Methyltransferases Detection.","authors":"Lin Hu, Honghua Hu, Fang Liu, Xingrong Li, Qinghua Wu, Xuhuai Fu, Diangeng Li, Xuan Wu, Rong Xiang, Rongzhong Huang, Xiaosong Li, Yi Li","doi":"10.1021/acssensors.6c02645","DOIUrl":"https://doi.org/10.1021/acssensors.6c02645","url":null,"abstract":"<p><p>DNA methyltransferases (MTases) are important epigenetic enzymes involved in DNA methylation regulation and have attracted considerable interest as analytical targets in disease-related epigenetic studies. However, traditional detection methods are constrained by laboratory-bound, sophisticated instrumentation and time-consuming protocols, while emerging approaches either depend on complex enzymatic amplifications or suffer from uncontrolled background signals, limiting their bioanalysis utility for precision-medicine-related applications. Inspired by the structure-controlled activation mechanism of CRISPR/Cas12a, we designed a branched split structure to regulate Cas12a for stepwise enzymatic activation and established the Cas12a-induced DNA MTase Sensor (CDMS), which enables highly sensitive MTase activity detection without the need for nucleic-acid amplification step. The core strategy lies in designing split-and-reconstituted Cas12a-activating DNA fragments with steric hindrance effects. This design significantly inhibits nonspecific Cas12a activation while efficiently restoring its activity upon removal of steric hindrance in response to DNA MTase activity. This method achieves ultra-low detection limit of 1.06 × 10-6 U/mL for M.SssI MTase and 7.41 × 10-8 U/mL for Dam MTase, exhibiting excellent specificity and robust applicability for MTase inhibitor screening. We also developed a palm-sized portable optical device, named iCas-Analyzer, to further explore the CDMS's potential for point-of-care testing (POCT), which can be easily operated by nonexpert end users. With device integration, the CDMS extends CRISPR-based diagnostics into the field of epigenetics, holding its potential for future decentralized MTase analysis and POCT-oriented assay development.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149147378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"PD-L1-Targeted and ROS-Responsive Nanoplatform for Simultaneous Chemotherapy and Dual-Modal Imaging of Treatment Response in Triple-Negative Breast Cancer.","authors":"Jingjing Hu, Chunting Wang, Yanhong Chen, Cuiying Zhu, Aoyu Liu, Dian Chen, Ling Zhan, Defan Yao, Dengbin Wang","doi":"10.1021/acssensors.6c03428","DOIUrl":"https://doi.org/10.1021/acssensors.6c03428","url":null,"abstract":"<p><p>The evaluation of treatment response in triple-negative breast cancer (TNBC) still primarily relies on tumor size and ex vivo analyses, which provide very limited information on how tumors dynamically respond during therapy. In fact, oxidative stress rapidly emerges early after drug treatment, while PD-L1 reflects an adaptive signal associated with tumors. However, these treatment-associated features are commonly assessed using static detection methods, thereby limiting longitudinal in vivo evaluation. To address this, we developed a theranostic nanoprobe named RAPP, which integrates DOX delivery, ROS-responsive near-infrared fluorescence (NIRF) imaging, and PD-L1-targeted magnetic resonance imaging (MRI). In this system, the fluorescence signal serves to report early oxidative stress in real time, while PD-L1 targeting contributes to tumor-directed MRI, enabling longitudinal assessment of both signals in a 4T1 TNBC model. During treatment, RAPP produced distinct changes in fluorescence and MR signals across different experimental groups, indicating the complementarity of these two imaging modalities in evaluating tumor response. Furthermore, DOX-loaded RAPP effectively inhibited tumor growth without exhibiting significant systemic toxicity. By integrating drug delivery with dual-modal imaging, RAPP offers a practical strategy for assessing tumor response in TNBC during treatment, rather than only after treatment completion.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149147193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-09DOI: 10.1021/acssensors.6c00595
Reddeppa Maddaka, Swetha Velpula, Justin Hutcheson, Hieu Pham Trung Nguyen
{"title":"III-Nitride Nanowire-Based Gas Sensors: From Material Design to Low-Temperature Sensing Performance.","authors":"Reddeppa Maddaka, Swetha Velpula, Justin Hutcheson, Hieu Pham Trung Nguyen","doi":"10.1021/acssensors.6c00595","DOIUrl":"https://doi.org/10.1021/acssensors.6c00595","url":null,"abstract":"<p><p>III-nitride nanowires (NWs), including gallium nitride (GaN) and its alloys, possess tunable direct band gaps, high electron mobility, and large breakdown voltages, making them highly promising for electronic and optoelectronic applications. In addition, their wide bandgap, efficient charge transport, mechanical robustness, and compatibility with CMOS technology make them strong candidates for high-performance sensing platforms. A gas sensor must meet certain qualities, such as low-temperature operation and high selectivity toward a particular gas, to prevent false signals caused by interference from other gases in the environment. Here, we review recent progress in room-temperature gas sensors based on III-nitride NW arrays, emphasizing defect-mediated gas adsorption, noble-metal decoration, integration with metal oxides and 2D materials, and light-activated sensing platforms. Detailed fabrication methods, sensing mechanisms, and strategies to enhance sensitivity, selectivity, and stability are discussed. Finally, current challenges and future design perspectives are highlighted to guide the development of practical, high-performance III-nitride NW gas sensors.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149134190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-09DOI: 10.1021/acssensors.6c02873
Alejandro Guzman-Landero, Ulrike Zanzen, Alexander Prange, Ramiro Marroquin-Garcia, Hanne Diliën, Bart van Grinsven, Rocio Arreguin-Campos
{"title":"Molecularly Imprinted Polymers as an Indirect Sensing Approach for Bacillus cereus Detection.","authors":"Alejandro Guzman-Landero, Ulrike Zanzen, Alexander Prange, Ramiro Marroquin-Garcia, Hanne Diliën, Bart van Grinsven, Rocio Arreguin-Campos","doi":"10.1021/acssensors.6c02873","DOIUrl":"https://doi.org/10.1021/acssensors.6c02873","url":null,"abstract":"<p><p>Bacterial endospores are a major challenge in food safety due to their high resistance and potential to cause spoilage and foodborne illness. Current detection methods can be labor-intensive and require specialized instrumentation. A low-cost, label-free thermal sensing platform for indirect detection of Bacillus cereus endospores using molecularly imprinted polymers (MIPs) targeting dipicolinic acid (DPA), a spore biomarker, was developed. The MIPs were synthesized by free-radical bulk polymerization, characterized by rebinding assays, and evaluated using the Heat Transfer Method (HTM). At 100 μM DPA, the MIPs exhibited an imprinting factor (IF) of 2.62 ± 0.18 (n = 3), demonstrating preferential DPA binding. Sensor performance was evaluated using aqueous DPA, structural analogues, and a complex food matrix. HTM measurements produced concentration-dependent responses, with limits of detection of 0.20 ± 0.15 μM in aqueous DPA solutions and 0.060 ± 0.0098 μM in unpretreated, spiked ultrahigh-temperature (UHT) milk. Terbium(III) fluorescence independently supported DPA retention by the MIPs. Real-time assays with germinated B. cereus suspensions at 105 colony-forming units (CFU) mL-1 enabled proof-of-concept monitoring of biologically released DPA under laboratory conditions. These findings highlight the potential of combining DPA-imprinted polymers with HTM for real-time and label-free indirect endospore detection.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149147205","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-09DOI: 10.1021/acssensors.6c02772
Feiyang Huang, Hao Wen, Zhiqing Gao, Guoqing Feng, Fengrong Liu, Ang Li, Yunrui Cao, Yuhang He, Qingjun Liu
{"title":"Functionalized Cellulose-Based Wearable Textile Detection from Insensible Sweat to a Multiparameter Biomarker in Sensible Sweat.","authors":"Feiyang Huang, Hao Wen, Zhiqing Gao, Guoqing Feng, Fengrong Liu, Ang Li, Yunrui Cao, Yuhang He, Qingjun Liu","doi":"10.1021/acssensors.6c02772","DOIUrl":"https://doi.org/10.1021/acssensors.6c02772","url":null,"abstract":"<p><p>Sweat is a key physiological fluid involved in regulating body temperature and maintaining hydration balance. Compared with sensible sweat, insensible sweat is continuously lost in the form of water vapor. Its dynamic variation is closely associated with skin barrier function and basic thermoregulatory activity. However, its low flux and diffuse distribution make reliable detection challenging. In addition, multiparameter analysis of biochemical components in sensible sweat is of significant importance for the evaluation of electrolyte balance and physiological status. In this work, a multiparameter textile platform based on functionalized cellulose materials was developed for monitoring and analyzing the sweating process. Cellulose was employed as the core material platform. By incorporating lithium chloride, humidity-responsive units were fabricated and further configured into a scalable fabric-integrable array, enabling the detection of subtle humidity variations induced by low-flux insensible sweat. As thermal load increased and the system transitioned to the sensible sweating stage, electrochemical sensing fibers were integrated. In combination with the sweat absorption and transport capability of cellulose yarns, the platform enabled the determination of key biochemical parameters in sensible sweat, including pH, sodium, potassium, and uric acid. On-body sweat analysis demonstrated that the textile system enables array-based monitoring of insensible sweat, during the transition to the sensible sweating stage, allowing simultaneous acquisition of multiple physiological parameters. This integrated capability provides a promising approach for continuous monitoring and evaluation of the human sweat rate, electrolyte balance, and related physiological conditions.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149134182","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-08DOI: 10.1021/acssensors.6c02932
Colby Hladun, Fung Kit Tang, Lawrence Tucker, Maheshwara Reddy Nadiveedhi, Jared Morse, Mahnoor Ali, Matthias Schmidt, Kaho Leung
{"title":"Leveraging In Situ Chlorination for Resolving Subcellular Chlorinative Stress.","authors":"Colby Hladun, Fung Kit Tang, Lawrence Tucker, Maheshwara Reddy Nadiveedhi, Jared Morse, Mahnoor Ali, Matthias Schmidt, Kaho Leung","doi":"10.1021/acssensors.6c02932","DOIUrl":"10.1021/acssensors.6c02932","url":null,"abstract":"<p><p>Hypochlorous acid (HOCl) is a key effector in host defense, serving as a potent mediator of the innate immune response. However, dysregulation of HOCl production contributes to tissue damage and chronic inflammation. Existing HOCl indicators are constrained by limited conjugation feasibility for subcellular targeting, narrow emission tunability, poor aqueous compatibility that requires high organic cosolvent content, or a lack of chlorination-specific sensing mechanisms. Here, we identify a previously unrecognized HOCl-mediated chlorination of 2,6-unsubstituted BODIPY scaffolds that produces pronounced increases in fluorescence emission wavelength, intensity, and fluorescence lifetime. We leveraged this reaction to develop a series of HOCl indicators, termed HOClsense dyes, for resolving chlorinative stress at the subcellular level. These HOClsense dyes provide switchable fluorescence turn-on and turn-off detection modes across tunable emission wavelengths, accompanied by redshifted responses. Strategic alkylation of the pyridyl group further enabled click-based functionalization, allowing the cellular distribution of HOClsense dyes to be tailored for subcellular imaging. Using HOClsense dyes, we resolved PAMP-induced extracellular and lysosomal HOCl and identified elevated basal HOCl levels and MPO-dependent hyperresponsive HOCl generation in Niemann-Pick disease cell models.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149134241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-08DOI: 10.1021/acssensors.6c01709
Wenying Tang, Chak Lam Jonathan Chan, Weiqi Zhang, Chen Wang, Yucheng Ding, Xiao Qiu, Zhu'an Wan, Zhesi Chen, Wenhao Ye, Zhiyong Fan
{"title":"An Artificial Olfactory Chip with Temperature Gradient Modulation for Food Waste Management.","authors":"Wenying Tang, Chak Lam Jonathan Chan, Weiqi Zhang, Chen Wang, Yucheng Ding, Xiao Qiu, Zhu'an Wan, Zhesi Chen, Wenhao Ye, Zhiyong Fan","doi":"10.1021/acssensors.6c01709","DOIUrl":"https://doi.org/10.1021/acssensors.6c01709","url":null,"abstract":"<p><p>Increasing demand for fuels driven by the scarcity of fossil resources necessitates the development of sustainable and innovative strategies for the energy industry. Food waste has emerged as a viable resource with diverse functional chemical components for energy harvesting. To enhance the efficiency and effectiveness of energy generation from food waste, classification and monitoring of food waste are crucial. Traditional vision recognition methods often face challenges due to waste overlaps and environmental constraints. In contrast, employing artificial olfactory chips (AOCs) holds promise in classifying food waste via gaseous information. However, bulky size, low sensitivity, limited classification ability, poor robustness, and lack of scalability hinder the implementation of AOCs into food waste management systems. Herein, we report a compact, monolithic AOC featuring a gas sensor array with temperature gradient modulation (TGM-Array), offering a promising platform for this application. Constructed on a porous anodized aluminum oxide (AAO) template with a high surface area, the TGM-Array chips demonstrate high sensitivity, detecting ethanol (EtOH) and hydrogen sulfide (H2S) at concentrations as low as 100 parts-per-billion (ppb). Leveraging the response patterns from sensing pixels at different operating temperatures, the TGM-Array chips exhibit effective discriminability. They categorized EtOH and H2S by type and concentration and distinguished selected food waste samples, such as beef and tomatoes, as well as their mixtures at different post-disposal durations under controlled experimental conditions. In addition, the TGM-Array chips maintain good robustness across four testing periods, identifying beef and tomato samples with varied post-disposal durations. Furthermore, their fabrication processes are photolithography-free and involve only three steps, rendering the TGM-Array chips cost-effective and well-suited for large-scale deployment in trash bins. These capabilities suggest the potential of TGM-Array chips for developing intelligent trash bin systems for improving food waste management efficiency and enhancing energy recovery processes.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149134231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-07DOI: 10.1021/acssensors.6c02316
Komal Jaswal,Victoria Aderonke Adekunle,Emma C. Tackman,Miriam Arak Freedman
{"title":"Dual Fluorophore System for Direct Measurement of Acidity of Aerosol Particles","authors":"Komal Jaswal,Victoria Aderonke Adekunle,Emma C. Tackman,Miriam Arak Freedman","doi":"10.1021/acssensors.6c02316","DOIUrl":"https://doi.org/10.1021/acssensors.6c02316","url":null,"abstract":"Abstract Atmospheric aerosol particles exhibit diverse physical and chemical properties, depending on their composition and source, which are challenging to measure directly due to their chemical complexity. Aerosol acidity is a key chemical property that has significant implications for climate, heterogeneous chemistry, and human health. Here, we present a novel approach to measure aerosol acidity using carbon dots (CDs) as in situ fluorophores. The method uses a dual fluorophore system consisting of pH-sensitive CDs and pH-insensitive CDs incorporated into model aerosol particles, with the pH-insensitive CDs serving as an internal standard. Fluorescence microscopy was used to construct a calibration curve by plotting the ratio of fluorescence intensities of droplets of fluorophore-containing standard solutions against pH. This calibration curve was used to determine the pH of model aerosol particles in the range of pH 1.5 to pH 3.5, demonstrating the efficacy of this approach for direct measurement of acidity. This approach holds significant potential to parameterize acidity in models, refine our understanding and predictions of aerosol properties such as heterogeneous chemistry, and aid in assessing the impact of aerosol particles on climate and health.","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":"405 1","pages":""},"PeriodicalIF":8.9,"publicationDate":"2026-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149091858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-07DOI: 10.1021/acssensors.6c02614
Lei Xia, Degen Chen, Yingying Deng, Changshun Sun, Lifang He, Jian Zhang, Kui Zhang
{"title":"Synergistic Effect of MOF-Derived Porous In2O3 and High-Entropy Alloy for Enhanced Room-Temperature Ammonia Sensing.","authors":"Lei Xia, Degen Chen, Yingying Deng, Changshun Sun, Lifang He, Jian Zhang, Kui Zhang","doi":"10.1021/acssensors.6c02614","DOIUrl":"https://doi.org/10.1021/acssensors.6c02614","url":null,"abstract":"<p><p>High-performance room-temperature ammonia (NH3) sensors are highly desirable for environmental monitoring and noninvasive chronic kidney disease diagnosis via breath analysis. Structurally advantageous, metal-organic framework (MOF)-derived metal oxides combined with low-cost, highly catalytic, nonprecious high-entropy alloy (HEA) nanoparticles show great potential for room-temperature gas sensing, yet such composites are rarely explored. Herein, FeCoNiMnCu HEA was rationally coupled with MOF-derived porous In2O3 to fabricate a HEA-modified In2O3 (HEA-IO) composite for room-temperature NH3 sensing. The optimized HEA-IO sensor delivered exceptional anti-interference selectivity and achieved a sensing response of 98.3 to 80 ppm NH3, almost twenty-fold that of pristine MOF-derived In2O3 and other control samples. Furthermore, it also featured rapid response/recovery speed, a low detection limit, humidity resistance, and excellent long-term stability. DFT calculations confirmed that FeCoNiMnCu HEA optimized surface electronic states, promoted NH3 enrichment and catalytic reactions, and accelerated interfacial charge transfer. Raman and TPD characterizations corroborated a unique interfacial synergistic interaction between HEA and MOF-derived In2O3, which was absent in conventional In2O3-based counterparts. The synergistic effect endowed HEA-IO with superior room-temperature NH3 sensing capabilities, enabling promising breath biomarker detection in clinical noninvasive diagnosis.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149126131","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS SensorsPub Date : 2026-10-07DOI: 10.1021/acssensors.6c02887
Jiayi He, Nathan A Rudman, Giulia Nisita, Ivan J Dmochowski
{"title":"Genetically Encoded Glucosensor for Multiplexed 129Xe NMR.","authors":"Jiayi He, Nathan A Rudman, Giulia Nisita, Ivan J Dmochowski","doi":"10.1021/acssensors.6c02887","DOIUrl":"https://doi.org/10.1021/acssensors.6c02887","url":null,"abstract":"<p><p>Hyperpolarized 129Xe NMR provides a non-optical platform for molecular, multiplexed detection. Genetically encoded protein reporters are attractive for biological sensing, but creating new 129Xe NMR sensors remains challenging because productive xenon binding is difficult to engineer into protein scaffolds. Here, we use glucose/galactose-binding protein (GGBP) as a ligand-responsive model protein to examine how molecular dynamics (MD)-derived cavity descriptors can guide the design of a xenon-binding protein sensor. Direct transfer of cavity-forming mutation from a related periplasmic binding protein (PBP) did not produce a well-resolved 129Xe response, motivating a broader analysis of candidate cavities by MD simulations. Among the GGBP variants examined, I24F/F90A combined an appropriate cavity volume, high pocket persistence, solvent exclusion, and high hydrophobicity density within the mutant series. The resulting glucosensor, GS, bound glucose with low-micromolar affinity and produced a distinct glucose-dependent 129Xe@GS hyper-CEST resonance shifted 30 ppm upfield from the aqueous xenon signal. This upfield resonance expands the chemical-shift window for genetically encoded protein-based 129Xe sensors to 125 ppm, highlighting an additional cavity-engineering route for modulating 129Xe NMR chemical shifts. GS enabled quantification of micromolar glucose concentrations in E. coli lysate, with results comparable to an enzymatic glucose assay. The upfield-shifted 129Xe NMR signal also allowed 129Xe@GS to be spectrally resolved from other PBP-based 129Xe sensors, enabling simultaneous detection of glucose-, ribose-, and maltose-responsive signals in a single hyper-CEST experiment. Finally, fusion of GS to the \"always-on\" 129Xe NMR contrast agent TEM-1 β-lactamase generated a ratiometric sensor. These results establish GGBP as a new scaffold for ligand-responsive 129Xe biosensing and introduce an MD-informed strategy for developing multiplexed and self-referenced genetically encoded 129Xe NMR sensors.</p>","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":10.9,"publicationDate":"2026-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149126072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}