Analytical Chemistry最新文献_第2页

Gas Sensing Based on Heterodyne Mach–Zehnder Interferometric Phase-Dispersion Spectroscopy 基于外差马赫-曾德干涉相色散光谱的气体传感

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-09-22 DOI: 10.1021/acs.analchem.6c04244

Yushuo Song, Shunda Qiao, Hanxu Ma, Ying He, Yufei Ma

{"title":"Gas Sensing Based on Heterodyne Mach–Zehnder Interferometric Phase-Dispersion Spectroscopy","authors":"Yushuo Song, Shunda Qiao, Hanxu Ma, Ying He, Yufei Ma","doi":"10.1021/acs.analchem.6c04244","DOIUrl":"10.1021/acs.analchem.6c04244","url":null,"abstract":"A novel gas sensing method based on heterodyne Mach–Zehnderinterferometric phase-dispersion spectroscopy (HMIPDS) was proposedfor the first time in this work. The proposed method employed a heterodyneMach–Zehnder interferometric configuration to realize gas sensingthrough cumulative optical phase shifts induced by refractive-indexvariations during laser-gas interaction. Specifically, the laser beamwas divided into a signal arm interacting with the gas medium anda reference arm frequency-shifted by an acousto-optic modulator (AOM)to generate heterodyne interference. The gas-induced optical phasevariation was then accurately extracted via quadrature demodulationof the interference signal, thereby enabling quantitative retrievalof gas concentration. Acetylene (C2H2) was usedfor experimental validation. Experimental results demonstrate theHMIPDS method achieved a detection limit of 708 ppb with an integrationtime of 500 s, corresponding to an approximately 11-fold improvementin detection sensitivity compared with the conventional WMS-2f detection system (8 ppm at 500 s). Furthermore, the lineardynamic detection range was extended from 0 to 2% for conventionalWMS-2f to 0–5%, corresponding to an approximately2.5-fold enhancement. The proposed HMIPDS method provides a high sensitivity,high stability, and wide dynamic range solution for gas sensing, showingstrong potential for high-concentration and ultrasensitive trace gasdetection applications.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29627–29638"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149019583","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}

引用次数: 0

Simple, Fast, and Ultrasensitive Fluorescent Detection of Peroxymonosulfate in Chemical, Enzymatic, and Cellular Systems 化学，酶和细胞系统中过氧单硫酸盐的简单，快速和超灵敏荧光检测

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-09-22 DOI: 10.1021/acs.analchem.6c03097

Radosław Podsiadły, Aleksandra Grzelakowska, Kumari Neelam, Radosław Michalski, Michał Rostkowski, Renata Smulik-Izydorczyk, Adam Sikora, Jacek Zielonka

{"title":"Simple, Fast, and Ultrasensitive Fluorescent Detection of Peroxymonosulfate in Chemical, Enzymatic, and Cellular Systems","authors":"Radosław Podsiadły, Aleksandra Grzelakowska, Kumari Neelam, Radosław Michalski, Michał Rostkowski, Renata Smulik-Izydorczyk, Adam Sikora, Jacek Zielonka","doi":"10.1021/acs.analchem.6c03097","DOIUrl":"10.1021/acs.analchem.6c03097","url":null,"abstract":"Peroxymonosulfate (PMS) is widely used in advanced oxidationprocesses(AOP) to remove organic contaminants from wastewater. A simple methodfor determining PMS is necessary to dose this oxidant effectivelyand use it adequately in AOP. Here, we report a simple, fast, andaccurate method for quantifying PMS using a pro-fluorescent boronateredox probe. In contrast to previously described methods, the proposedassay does not require activation of PMS by a low-valent transitionmetal. Based on the determined second-order rate constant of the directoxidation reaction of the coumarin-7-boronic acid probe with PMS (2k = 1.1 × 104 M–1 s–1), we have developed a method based on high-performanceliquid chromatography coupled with fluorescence detection (HPLC-FLD)for ultrasensitive detection of PMS with a detection limit of 5 nMand a quantification limit of 40 nM. The HPLC-FLD method enables detectionof PMS in real water samples, confirming its practical usefulness.The reported method enables the determination of PMS over a wide pHrange and in the presence of common ions. It also enables trackingof PMS formation in sulfite solutions. Using activated RAW 264.7 macrophages,we demonstrate the feasibility of real-time monitoring of PMS productionusing a plate reader with fluorescence detection, along with appropriatecontrol experiments.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29541–29552"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149019594","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}

引用次数: 0

Correction to “Low-Perturbation Whole-Blood CD11b Measurement by Biomimetic Acoustofluidic Preprocessing and Wash-Free Microsphere-Competition Readout” 对“低摄动全血cd11b测量的仿生声流预处理和免洗微球竞争读数”的修正

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-10-06 DOI: 10.1021/acs.analchem.6c06557

Yifan Wang, Yihan Ma, Xiaotong Si, Chen Yang, Mengyuan Xu, Keyu Chen, Rui You, Fubing Wang, Fuling Zhou, Yi Yang

引用次数: 0

Electrophoresis-Correlative diaPASEF Improves Low-Input and Single-Cell CE−ESI−MS Proteomics by Targeting Peptide-Enriched Acquisition Space 电泳-相关diaPASEF提高低输入和单细胞CE - ESI - MS蛋白质组学靶向肽富集获取空间

IF 7.4 1区化学

Analytical Chemistry Pub Date : 2026-10-20 DOI: 10.1021/acs.analchem.6c03247

Fei Zhou,Bowen Shen,Nicolas Drouin,Peter Nemes

{"title":"Electrophoresis-Correlative diaPASEF Improves Low-Input and Single-Cell CE−ESI−MS Proteomics by Targeting Peptide-Enriched Acquisition Space","authors":"Fei Zhou,Bowen Shen,Nicolas Drouin,Peter Nemes","doi":"10.1021/acs.analchem.6c03247","DOIUrl":"https://doi.org/10.1021/acs.analchem.6c03247","url":null,"abstract":"In low-input proteomics, proteome coverage is often limited not only by sample amount but also by inefficient tandem mass spectrometry (MS) sampling of peptide-rich acquisition space. During capillary electrophoresis (CE)−electrospray ionization (ESI)−MS analysis, electrophoretically separated peptides occupy reproducible, correlated regions of migration time, precursor mass-to-charge ratio (m/z), and ion mobility (IM), creating an opportunity to direct data-independent acquisition (DIA) toward peptide-enriched regions. Here, we introduce electrophoresis-correlative diaPASEF (Eco-diaPASEF), a CE-informed implementation of diaPASEF that uses migration time-correlated m/z and IM information to define complementary acquisition segments while maintaining the reference cycle time. By restricting IM scan range, precursor m/z space, and DIA isolation width, Eco-diaPASEF focuses tandem MS acquisition on peptide-enriched regions. In low-input HeLa digest samples, Eco-diaPASEF increased protein-group (PG) identifications by up to ∼65% relative to conventional diaPASEF, cumulatively identifying up to 1163 PGs across 4 technical replicates from 200 pg of digest measured per run on a timsTOF Pro mass spectrometer. Applied to single animal-cap cells from stage-9 Xenopus laevis embryos, Eco-diaPASEF cumulatively quantified 1108 PGs across 3 cells using ∼200 pg of injected digest per cell. The method provided ∼2.6−2.7-fold greater proteome coverage across replicate criteria while maintaining comparable biological-replicate precision. These results establish Eco-diaPASEF as a CE-guided acquisition strategy for improving proteome coverage in low-input and single-cell CE−ESI−MS proteomics on IM-enabled mass spectrometers.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"56 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221672","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}

引用次数: 0

Monitoring of Doxorubicin-Mediated Mitochondria–Lysosome Crosstalk by In Situ Raman Spectroscopy: The Molecular Mechanism during Apoptosis and Implications for Cardiotoxicity 原位拉曼光谱监测阿霉素介导的线粒体-溶酶体串扰：凋亡过程中的分子机制及其对心脏毒性的影响

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-09-19 DOI: 10.1021/acs.analchem.6c03083

Li Song, Zeyi Wang, Yaqiang Chen, Wei Li, Jinyu Zhu, Linjun Cai, Xiao Xia Han

{"title":"Monitoring of Doxorubicin-Mediated Mitochondria–Lysosome Crosstalk by In Situ Raman Spectroscopy: The Molecular Mechanism during Apoptosis and Implications for Cardiotoxicity","authors":"Li Song, Zeyi Wang, Yaqiang Chen, Wei Li, Jinyu Zhu, Linjun Cai, Xiao Xia Han","doi":"10.1021/acs.analchem.6c03083","DOIUrl":"10.1021/acs.analchem.6c03083","url":null,"abstract":"Doxorubicin is among the most commonly used chemotherapeuticagentsfor a wide range of cancers. Despite its effectiveness, it poses long-termcardiotoxic risks that elevate mortality among cancer patients. Inthis study, doxorubicin-mediated mitochondria–lysosome crosstalkin apoptosis and the underlying molecular mechanism are explored.The interactions between organelle-specific phospholipids and doxorubicin,and their structural basis, are investigated using comprehensive spectroscopyand molecular dynamics simulations. Our results by in situ Raman spectroscopyreveal that doxorubicin can directly trigger mitochondrial and lysosomalpermeabilization, facilitating the release of key apoptotic modulators.Besides phospholipids, doxorubicin is found to interact with cytochromec and boost its proapoptotic effect. Further study indicates thatprotecting cardiolipin is sufficient to inhibit doxorubicin-induceddisruption of CL conformation. Accordingly, the molecular mechanismunderlying doxorubicin-mediated mitochondria–lysosome crosstalkin apoptosis is proposed, paving the way for the rational design ofcardioprotective agents targeting mitochondria and lysosomes for cancerpatients.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29529–29540"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149003345","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}

引用次数: 0

Drug Metabolism and Drug–Drug Interactions Studied Using High-Throughput Ambient Mass Spectrometry 高通量环境质谱法研究药物代谢和药物-药物相互作用

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-09-23 DOI: 10.1021/acs.analchem.6c04039

Mousumi Saha, Nari Talaty, Nicolás M. Morato, Mahdiyeh Shahi, Christopher J. Welch, Yunfei Feng, Christina R. Ferreira, David Wagner, Andreas Kaerner, Robert Graham Cooks

{"title":"Drug Metabolism and Drug–Drug Interactions Studied Using High-Throughput Ambient Mass Spectrometry","authors":"Mousumi Saha, Nari Talaty, Nicolás M. Morato, Mahdiyeh Shahi, Christopher J. Welch, Yunfei Feng, Christina R. Ferreira, David Wagner, Andreas Kaerner, Robert Graham Cooks","doi":"10.1021/acs.analchem.6c04039","DOIUrl":"10.1021/acs.analchem.6c04039","url":null,"abstract":"Rapid characterization of drug metabolism and drug–druginteractions (DDIs) with quantitative rigor remains challenging dueto complex samples and labor-intensive workflows. We present a high-throughput(HT) ambient mass spectrometry (MS) approach using desorption electrosprayionization (DESI) that unites three complementary capabilities availableusing two different mass spectrometers: (A) untargeted screening tosurvey the metabolic space, (B) product ion scan tandem MS to characterizenovel metabolites, and (C) multiple reaction monitoring (MRM) to deliverprecise quantitation of substrates and metabolites. Applying the methodologyto diverse in vitro systems, we track substrate depletion,metabolite formation, and DDIs with quantitative accuracy (error <9%for MRM quantitation) validated against liquid chromatography (LC)MS. The method operates two to three orders of magnitude faster (1s/sample) than traditional LC–MS methods. This allows hundredsof substrate–enzyme–DDI combinations to be followedin time-resolved fashion using ∼4 × 104 individualDESI-MS measurements, acquired at a rate of >3,000 measurements/hour.By combining HT screening and targeted monitoring, the DESI-MS approachdelivers metabolic discovery with robust quantitation, overcominga central bottleneck in large-scale drug metabolism studies. Thisinvestigation aims to provide a high-throughput analytical approachwith broad utility in drug discovery, toxicology, and precision medicineworkflows.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29604–29615"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149019599","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}

引用次数: 0

Quantifying Viscosity of Semisolid Submicrometer and Nanosized Individual Aerosols Using Atomic Force Microscopy with Varying Cantilever Velocity 半固体亚微米和纳米尺度单个气溶胶粘度的原子力显微镜定量研究

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-10-01 DOI: 10.1021/acs.analchem.6c02398

Ruwini A. Weerasinghe, Chamika K. Madawala, Alexei V. Tivanski

{"title":"Quantifying Viscosity of Semisolid Submicrometer and Nanosized Individual Aerosols Using Atomic Force Microscopy with Varying Cantilever Velocity","authors":"Ruwini A. Weerasinghe, Chamika K. Madawala, Alexei V. Tivanski","doi":"10.1021/acs.analchem.6c02398","DOIUrl":"10.1021/acs.analchem.6c02398","url":null,"abstract":"Atmospheric aerosol viscosity governs diffusion timescales ofreactive gases into aerosols, thereby influencing gas uptake, heterogeneouschemistry, and atmospheric aging. However, direct viscosity measurementsof individual semisolid aerosols have been restricted to a relativelyhigh viscosity range and submicrometer size. Herein, a new atomicforce microscopy-based nanorheological experimental approach is presentedthat utilizes force–indentation distance and indentation ratemeasurements at varying cantilever velocities to quantify the viscosityof individual semisolid aerosols as a function of relative humidity(RH). Using glucose and sucrose as model systems, we demonstrate accuratesingle particle viscosity quantification spanning ∼102·5 to 107 Pa s under atmospherically relevantRH conditions. We establish an iterative cantilever velocity frameworkthat enables the determination of the appropriate velocity requiredfor accurate viscosity characterization of aerosols without priorknowledge of their composition. Additionally, we demonstrate thatthe method enables accurate viscosity quantification of individualsub-100 nm aerosols. We anticipate that future viscosity measurementsof atmospheric aerosols from various sources at different RHs willprovide valuable insights into aerosol viscosity variability and revealthe extent of particle-to-particle heterogeneity.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29471–29480"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149091825","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}

引用次数: 0

SPRINT-Cas12a: A Single Leading-Hairpin Probe-Enabled Dual-Pathway Activation Strategy for Sensitive miRNA Detection SPRINT-Cas12a：用于敏感miRNA检测的单导发夹探针双通路激活策略

IF 7.4 1区化学

Analytical Chemistry Pub Date : 2026-10-20 DOI: 10.1021/acs.analchem.6c06144

Qian Li,Junda Liu,Mo Xue,Jiahao Wang,Youfei Zhang,Luzhen Huai,Xingzhou Zhu,Xiaoyan Liu,Jie Wang

{"title":"SPRINT-Cas12a: A Single Leading-Hairpin Probe-Enabled Dual-Pathway Activation Strategy for Sensitive miRNA Detection","authors":"Qian Li,Junda Liu,Mo Xue,Jiahao Wang,Youfei Zhang,Luzhen Huai,Xingzhou Zhu,Xiaoyan Liu,Jie Wang","doi":"10.1021/acs.analchem.6c06144","DOIUrl":"https://doi.org/10.1021/acs.analchem.6c06144","url":null,"abstract":"CRISPR−Cas12a possesses robust trans-cleavage activity and has emerged as a powerful tool for nucleic acid detection. However, the efficient integration of Cas12a trans-cleavage with direct miRNA recognition and signal amplification remains challenging, particularly because conventional assay designs often rely on multiple probes or a single activation pathway. Here, we developed SPRINT-Cas12a, a single-probe RNA-triggered integrated trans-cleavage platform for sensitive miRNA detection. In this design, a leading hairpin probe (L-HP) specifically recognizes the target miRNA and initiates polymerase-mediated cascade isothermal amplification, synchronously generating two structurally distinct Cas12a-activating products: an RNA−DNA complex and a long-chain micro-hairpin probe (L-MHP). These products provide complementary activation pathways for Cas12a, thereby promoting reporter cleavage and fluorescence signal generation. The single-probe architecture simplifies probe design while enabling synergistic signal amplification through dual-pathway Cas12a activation. The platform was successfully adapted for the detection of miR-21 and miR-122, two miRNAs with distinct sequences, demonstrating its modularity and sequence versatility. Using miR-192 as a hepatocellular carcinoma-associated target, the method achieved sensitive detection in serum samples and distinguished patients with hepatocellular carcinoma from healthy controls, with results consistent with stem-loop quantitative reverse-transcription PCR. In addition, the platform was coupled with a lateral flow assay to provide an instrument-free visual readout. Depending on the amplification and readout format, the assay could be completed within 90 min to 120 min using simple laboratory equipment. Overall, SPRINT-Cas12a provides a single-probe, dual-pathway strategy for Cas12a-mediated miRNA detection and shows promise for portable molecular diagnostics and point-of-care testing.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"13 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221674","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}

引用次数: 0

Physics-Informed Machine Learning Overcomes Particle-Size Nonlinearities That Confound the Raman Spectrometry of Granular Carbonaceous Materials 机器学习克服了颗粒尺寸的非线性，这种非线性混淆了颗粒碳材料的拉曼光谱

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-09-25 DOI: 10.1021/acs.analchem.6c02307

Ginger W. Brown, Natalia V. Solomatova, Matthew D. Kowal, Miayan Larose, Calum Cole, Nicola Jurinovic, Sadegh Shokatian, Edward Grant

{"title":"Physics-Informed Machine Learning Overcomes Particle-Size Nonlinearities That Confound the Raman Spectrometry of Granular Carbonaceous Materials","authors":"Ginger W. Brown, Natalia V. Solomatova, Matthew D. Kowal, Miayan Larose, Calum Cole, Nicola Jurinovic, Sadegh Shokatian, Edward Grant","doi":"10.1021/acs.analchem.6c02307","DOIUrl":"10.1021/acs.analchem.6c02307","url":null,"abstract":"Raman spectroscopy supports multivariate regression modelsthatcan offer a rapid and nondestructive approach to quantifying the compositionof granular materials. Soils constitute a class of such materialsfor which chemical analysis provides vital information for agricultureand environmental stewardship. However, soil samples typically presenthighly heterogeneous mixtures of varying particle size and diverseoptical properties owing to light-absorbing constituents. Althoughparticle-size effects on Raman spectrometry in scattering media havelong been considered in the context of pharmaceutical analytical technology,the quantitative influence of particle size in opaque, absorbing materialsremains poorly understood. Here, we investigate the effect of particlesize and light absorption in controlled mixture systems that mimicsoils. These mixtures, containing particles of varying sizes and opticalproperties, isolate the ways in which fluorescence, granularity andlight absorption affect the accuracy of machine learning regressionmodels. Using a dual-laser Raman system, we collected both conventionalRaman and Shifted-Excitation Raman Difference Spectroscopy (SERDS)spectra, showing that light-absorbing compounds attenuate Raman intensitiesnonlinearly to an extent that varies with particle size. Multivariateregression models link this behavior to optical extinction governedby the surface area of the absorbing grains. We account for this behaviorin terms of a simple physical model that incorporates the physicsof light scattering and absorption in heterogeneous systems of varyingparticle size. This hybrid, physics-informed machine learning frameworkoutperforms purely data-driven approaches in predicting sample compositionfrom Raman spectra, highlighting the importance of physical characteristicsin the analysis of light-absorbing granular materials.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29459–29470"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149049911","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}

引用次数: 0

Permeability-Based Measurement of Nanobody Binding to Nuclear Pore Complexes by Transient Scanning Electrochemical Microscopy 基于渗透率的纳米体与核孔复合物结合的瞬态扫描电化学显微镜测量

IF 7.3 1区化学

Analytical Chemistry Pub Date : 2026-10-20 Epub Date: 2026-09-24 DOI: 10.1021/acs.analchem.6c04319

Moghitha Parandhaman, Siao-Han Huang, Kathryn G. Dzurik, W. Seth Childers, Shigeru Amemiya

{"title":"Permeability-Based Measurement of Nanobody Binding to Nuclear Pore Complexes by Transient Scanning Electrochemical Microscopy","authors":"Moghitha Parandhaman, Siao-Han Huang, Kathryn G. Dzurik, W. Seth Childers, Shigeru Amemiya","doi":"10.1021/acs.analchem.6c04319","DOIUrl":"10.1021/acs.analchem.6c04319","url":null,"abstract":"Nucleocytoplasmic macromolecular transport through thenuclearpore complex (NPC) is mediated selectively and efficiently by protein–proteininteractions, thereby regulating gene expression and the cell cycle.The NPC transport barrier is formed by biomolecular condensates ofnucleoporins (nups) rich in hydrophobic phenylalanine–glycine(FG) repeats. Herein, we develop the transient mode of scanning electrochemicalmicroscopy (SECM) to measure NPC interactions with a nanobody proteintargeting the non-FG site of the crucial barrier-forming FG-rich nup,Nup98. The small nanobody, which was recombinantly expressed and purified,diffuses into the NPC and partially blocks the transport of an FG-bindingpolycationic peptide, protamine. The lowered permeability is measuredmore sensitively by transient SECM than by the traditional steady-statecounterpart under our experimental conditions. A dissociation equilibriumconstant between the nanobody and Nup98 in the authentic NPC is determinedfrom the dependence of polypeptide permeability on nanobody concentration.The resulting dissociation constant of 7.7 nM is significantly higherthan that of 1.7 nM measured for isolated Nup98 with the nanobodyattached to the optical sensor surface in biolayer interferometry.The higher dissociation constant of nanobody–Nup98 interactionsin the NPC is consistent with the enthalpically and entropically unfavorablepartitioning of the nanobody into the condensate of hydrophobic anddisordered nups. The powerful SECM method will be applicable to investigatea variety of NPC-binding proteins with the authentic NPC, therebyadvancing our understanding of protein–NPC interactions andfunctions.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29639–29647"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149049987","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}

引用次数: 0