Kenneth R Durbin,Matthew T Robey,Joseph B Greer,Ryan T Fellers,Aaron O Bailey
{"title":"Fast and Accurate Charge State Deconvolution of Protein Mass Spectra.","authors":"Kenneth R Durbin,Matthew T Robey,Joseph B Greer,Ryan T Fellers,Aaron O Bailey","doi":"10.1021/acs.analchem.5c00288","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00288","url":null,"abstract":"Charge state deconvolution is essential for efficient and effective protein mass spectrometry analysis. High-quality mass profiling is necessary to determine which proteoforms are present in protein samples and their relative abundances. In the pursuit of a well-rounded deconvolution solution, we detail an iterative charge state deconvolution algorithm named kDecon that has been tuned to provide high accuracy in its mass results while also delivering superb sensitivity toward lower abundance proteoforms in complex spectra. Here, the performance of kDecon as a mass determination algorithm for both targeted antibody and high-throughput proteomics analysis was benchmarked against existing deconvolution solutions. While the different deconvolution routines all proved robust for detecting the highest abundance protein species, kDecon ultimately showcased best-in-class precision for lower abundance proteoform mass profiling. Furthermore, kDecon results had up to 7-fold fewer false positives and simultaneously exhibited at least 20-fold speed improvements over the other algorithms. Overall, these deconvolution advances will contribute to enabling both routine and thorough intact mass profiling studies for biotherapeutics as well as improving the proteome coverage of top-down proteomics experiments.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"2 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568595","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":"Exploring Meso-Six-Membered-N-Heterocycle BODIPY Rotors as AIE Probes for Fluorescence Imaging Subcellular Viscosity in Organelles.","authors":"Jin-Cheng Li,Yingmei Cao,Wei-Lin Lu,Yu-Xin Tang,Xi-Xi Zheng,Wen-Jing Shi,Liyao Zheng,Jin-Wu Yan,Dongxue Han,Li Niu","doi":"10.1021/acs.analchem.4c06607","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06607","url":null,"abstract":"Aggregation-induced emission (AIE)-type small-molecular fluorescent rotors have attracted wide interest due to their inherent excellent properties in bioimaging viscosity, which can potentially reveal the relationships between microviscosity and related diseases. Although many fluorescent probes have been designed, easy-to-prepare and multifunctional AIE rotors remain a hot topic and less reported. Herein, four easy-to-prepare and water-soluble meso-pyrimidine/pyridine BODIPY-based multifunctional fluorescent probes were rationally designed to investigate the effects of the number and position of nitrogen in six-membered-N-heterocycles on viscosity, AIE, and viscosity imaging. Luckily, the meso-o-pyrimidine-based probe 1 displayed significant fluorescence enhancements at 525 nm with gradually increasing viscosity from water to glycerol, which also showed obvious AIE property in an acetonitrile/water mixture due to two nitrogen at the ortho positions of pyrimidine. Meso-o-pyridine-based 2 exhibited no viscosity-enhanced emissions, but obvious AIE enhancements appeared. Comparatively, the meso-m-pyridine-based probe 3 showed no viscosity- and aggregation-induced emission enhancements, while the meso-p-pyridine-based one 4 displayed obvious viscosity response without AIE character. Furthermore, 1 and 4 were applied to cellular imaging, which displayed lysosomal and mitochondrial localizations in HeLa cells, respectively, and successfully monitored viscosity variations induced by the addition of lipopolysaccharide (LPS) or monensin. In summary, four meso-pyrimidine and meso-pyridine-BODIPY multifunctional fluorescent rotors have been prepared using easy synthesis, in which meso-o-pyrimidine and meso-o-pyridine ones showed improved AIE properties in water, and meso-o-pyrimidine and meso-p-pyridine ones showed good subcellular localization and viscosity changes in HeLa cells. This work can provide an easy-to-prepare and useful strategy for multifunctional fluorescent probes with enhanced AIE property for imaging viscosity and other applications.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"151 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568635","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}
Jialing Zhong,Yong Chen,Yueyu Dong,Chuanghao Guo,Yizhen Liu
{"title":"SPARC: An Orthogonal Cas12a/Cas13a Dual-Channel CRISPR Platform for Reliable SNV Identification and Mutation Confirmation.","authors":"Jialing Zhong,Yong Chen,Yueyu Dong,Chuanghao Guo,Yizhen Liu","doi":"10.1021/acs.analchem.5c02141","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c02141","url":null,"abstract":"Rapid and reliable detection of single nucleotide variants (SNVs) is essential for accurate pathogen diagnostics, genetic mutation screening, and personalized medicine. However, existing CRISPR-based nucleic acid detection platforms frequently suffer from ambiguous signal interpretation, specificity limitations, and complex assay workflows. Herein, we introduce SPARC (specific and precise mutation recognition with Cas12a/Cas13a), a novel orthogonal dual-channel CRISPR assay that significantly enhances the SNV detection reliability. SPARC integrates Acidaminococcus sp. Cas12a (AsCas12a), which specifically detects a conserved region as an internal reference, with our recently identified DNA-activated Leptotrichia buccalis Cas13a (LbuCas13a), which exhibits exceptionally high intrinsic SNV specificity without requiring engineered crRNA mismatches. The orthogonal design uniquely resolves the common diagnostic ambiguity between genuine SNVs and target absence. Combined with recombinase polymerase amplification (RPA) and T7 exonuclease digestion, the SPARC platform achieved a sensitivity as low as 1 aM. We demonstrated the platform's robust clinical applicability through successful detection and accurate differentiation of hepatitis B virus (HBV) and clinically significant YMDD resistance mutations. This work presents an innovative and versatile CRISPR-based solution, highlighting substantial potential for advancing clinical diagnostics and precision medicine.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"2 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568633","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}
Chi Soo Park,Chulmin Moon,Han Seul Lee,Kyuran Kim,Haeun Byeon,Daeun Eom,Siwon Kim,Seojeong Lee,Ha Hyung Kim
{"title":"Site-Specific Identification of α2,6 and/or α2,3 Sialyl-Linkage Isomers in CTLA4-Igs Using Nano-LC-Quadrupole-Orbitrap-MS/MS.","authors":"Chi Soo Park,Chulmin Moon,Han Seul Lee,Kyuran Kim,Haeun Byeon,Daeun Eom,Siwon Kim,Seojeong Lee,Ha Hyung Kim","doi":"10.1021/acs.analchem.5c01699","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01699","url":null,"abstract":"Cytotoxic T-lymphocyte-associated antigen-4 (CTLA4)-Ig is a fusion protein used to treat autoimmune diseases. In glycoproteins, sialic acids are linked to galactose via either α2,6 or α2,3 linkages, with the latter being more rigid and crucial for extending the serum half-life of biotherapeutics. However, site-specific identification of α2,6 and/or α2,3 linkages in glycoproteins remains a significant analytical challenge. In this study, we performed site-specific N-glycan analysis with sialyl-linkage isomers in a CTLA4-Ig mutant (CTLA4-IgMT) produced in Chinese hamster ovary (CHO) cells co-overexpressing N-acetylglucosaminyltransferase-IV and α2,6-sialyltransferase. Since CHO cells produce α2,3-linked glycoproteins, wild-type CTLA4-Ig (CTLA4-IgWT) was used as a control. Following Pronase digestion, N-glycopeptides were enriched and analyzed using nano-liquid chromatography-quadrupole-Orbitrap-tandem mass spectrometry. Retention time analysis revealed that α2,6-linked N-glycopeptides exhibit lower hydrophobicity than their α2,3-linked counterparts. Fragmentation analysis showed that fragment ion intensity ratios ([sialic acid]+/[N-acetylglucosamine]+) are lower for α2,6 linkages. To improve the accuracy, multiple linear regression was applied to adjust fragment ion intensities, deriving compensation coefficients for N-acetylglucosamine, galactose, and sialic acid. Using this approach, structural alterations in CTLA4-IgMT were identified, including 6 additionally generated, 11 partially converted, and 6 completely converted N-glycans, yielding an overall α2,6 linkage proportion of 56.7%. Notably, α2,6 linkages were identified at N76 and N108 in the CTLA4 region but were absent at N207 in the Fc region. This study is the first to determine α2,6 and/or α2,3 linkages at three N-glycosylation sites in CTLA4-Ig, providing a strategy for distinguishing sialyl-linkage isomers.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"685 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568632","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":"High-Flux Chip for Sensitive Profiling of Small Extracellular Vesicle Proteins by Cas9/Switch-sgRNA Complex-Mediated Proximity Cleavage Assay.","authors":"Xiaoli Yao,Haixia Shi,Shoukun Wu,Xiang Xue,Zhengyu Xu,Zichuan Ping,Xiaowen Qiu,Qingfeng Hu,Guojun Wei","doi":"10.1021/acs.analchem.5c01467","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01467","url":null,"abstract":"Small extracellular vesicles (sEVs) play pivotal roles in modulating the pathological processes of various diseases and have emerged as promising biomarkers for disease diagnosis, including acute spinal cord injury and cancers. This is attributed to their ability to transport multiple proteins that reflect the molecular signatures of their parent cells. The evaluation of surface proteins presents a robust strategy for identifying a comprehensive set of biomarkers. In this study, we developed a high-throughput device capable of characterizing surface proteins on intact sEVs. Our approach employs CD63 antibodies immobilized on a 96-well plate and a CD9 aptamer integrated into switch-sgRNA, facilitating the efficient capture of intact sEVs and enabling subsequent surface protein profiling. The system utilizes a proximity cleavage assay mediated by the Cas9-nickase/switch-sgRNA complex and an identity probe, combined with DNA polymerase-assisted chain extension and displacement, to achieve highly specific and precise identification of target proteins. The DNA polymerase-mediated chain extension and displacement mechanism within the wells generates multiple G-rich sequences, which facilitate Thioflavin T (ThT)-based label-free signal amplification. This innovative design allows the high-throughput chip to profile the surface protein EpCAM on intact sEVs with exceptional sensitivity, achieving a remarkably low detection limit of 3.5 particles/μL. Moreover, the chip has been successfully applied to identify surface markers including EpCAM, PTK7, PDGF, and PSMA on sEVs derived from various biological samples, demonstrating its significant potential for high-throughput biomarker discovery and analysis.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"8 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568636","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}
Arno Krause,Gabriel Giardina,Laszlo Papp,David Haberl,Clemens P Spielvogel,Richard D Walton,James Marchant,Nestor Pallares-Lupon,Kanchan Kulkarni,Xu Li,David L Vasquez,Jürgen Popp,Iwan W Schie,Wolfgang Drexler,Marco Andreana,Angelika Unterhuber
{"title":"Multimodal Optical Imaging Combined with Radiomic Analysis for Fibrotic Cardiac Tissue Investigation.","authors":"Arno Krause,Gabriel Giardina,Laszlo Papp,David Haberl,Clemens P Spielvogel,Richard D Walton,James Marchant,Nestor Pallares-Lupon,Kanchan Kulkarni,Xu Li,David L Vasquez,Jürgen Popp,Iwan W Schie,Wolfgang Drexler,Marco Andreana,Angelika Unterhuber","doi":"10.1021/acs.analchem.5c01510","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01510","url":null,"abstract":"Understanding the process of fibrotic scarring of the myocardium is critical for the diagnosis and risk stratification of life-threatening cardiac dysfunction. Complex changes in structure, composition, and conductivity occurring at different stages of fibrogenesis diversify the biomedical characteristics of the myocardium. We present a multimodal optical imaging approach including cardiac optical mapping (COM), optical coherence tomography (OCT), multiphoton microscopy (MPM), and line scan Raman microspectroscopy (LSRM) for multiparametric assessment of the myocardium with radiomic analysis to link electrophysiologic, morphologic, functional, and molecular changes in ischemic cardiac tissue and validate our results with histology. COM is used to map the electrical behavior across myocardial tissue. Second harmonic generation and two-photon excitation fluorescence imaging as MPM techniques provide additional unique contrast of collagen, the extracellular matrix, and cardiac cells, such as cardiomyocytes playing a critical role in cardiac fibrosis. Our machine learning model based on radiomic features extracted from MPM data addresses the need for automated fast high-throughput classification between healthy and pathologic cardiac tissues and achieved an accuracy of 0.99. In addition, LSRM assesses the molecular contrast and is used to evaluate the development stage of fibrotic scarring and multiclass classification by utilizing partial least-squares discriminant analysis, achieving sensitivity and specificity values of 0.94. OCT is used for fast navigation through the sample, for intermodal referencing, and easy coregistration between the complementary imaging techniques operating at different fields of view and resolutions ranging from cm2 down to μm2.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"4 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144566167","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":"One-Step Strategy to Synthesize Sulfonic Acid-Functionalized Silica Chromatographic Stationary Phases for Separation of Rare-Earth Elements.","authors":"Yuqing Wei,Yali Yang,Chao Zhong,Chen Shen,Shuaishuai Wang,Jia Chen,Hongdeng Qiu","doi":"10.1021/acs.analchem.5c01206","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01206","url":null,"abstract":"Separation and analysis of rare-earth elements (REEs) with impurities are challenging processes. Ion-exchange chromatography has emerged as a valuable technology for rare-earth separation, and developing a simple and efficient preparation method for stationary ion-exchange chromatography remains a hot topic. In this study, a sulfonic acid-modified silica stationary phase was synthesized through a facile one-step strategy, where 1,3-propanesultone reacted with the silanol group. The stationary phase demonstrated effective cation-exchange chromatographic performance, achieving baseline separation of 15 REEs with resolution (RS) ranging from 1.9 to 6.8 for adjacent lanthanides. Excellent separation capability was also observed for impurity ions, including Fe, U, and Th, from the REEs. The stationary phase demonstrated an enhanced separation performance for REEs compared to conventional bonded phases. In addition, the columns exhibited excellent repeatability and stability. The separation performance was systematically evaluated across particle sizes of 3, 5, and 10 μm (with surface coverages of 0.89, 0.70, and 0.58 μmol/m2, respectively), demonstrating the universality of the preparation method. These findings support its potential application for preparative-scale stationary phases to separate REEs.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"97 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144566166","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":"Real-Time Fluorescence Aptasensor Based on Programmable Switch-Coupled Exponential Amplification Transduction for Ultrasensitive and Specific Vibrio parahemolyticus Determination.","authors":"Yinglong He,Wenxuan Li,Jiaqi Zhao,Xueyu Yang,Jiaqi Chen,Chen Ju,Wenjing Jin,Xiaogai Hou,Xinran Xiang,Shenghang Zhang","doi":"10.1021/acs.analchem.5c01104","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01104","url":null,"abstract":"The accurate and early detection of Vibrio parahemolyticus (V. parahemolyticus) is crucial for ensuring food safety and protecting consumer health. However, current detection methods face challenges in preventing the rapid transmission of V. parahemolyticus due to insufficient sensitivity and limited coverage. Herein, an aptamer-based real-time fluorescence biosensor was developed, termed CAPANA (a catalytic nucleic acid exponential cascade amplification network-powered magnetic aptasensor) for simple and ultrafast sensitive diagnosis of V. parahemolyticus. By integrating the programmable truncated aptamer and the EXPAR-TtAgo cascade reaction, the CAPANA system dexterously converts the epitope recognition events of V. parahemolyticus into fluorescence signals. The CAPANA system can not only address the low sensitivity of the single aptasensor transduction and the interference from nonessential bases in amplicons but also maintain dynamic balance between repeated regeneration of target and exponential signal amplification. Moreover, considering the potential variations in reaction conditions among multiple enzymes and the possible impacts of their interactions on reaction efficiency and specificity, engineering improvements in the CAPANA system can optimize coordination and complexity among multiple enzymes. The CAPANA system demonstrated a 60.61-fold higher detection efficacy for V. parahemolyticus than the TtAgo-based cleavage method. The detection limit achieved is 100 CFU/mL, and the rapid results take only 40 min. This system was confirmed by identifying contaminated aquatic products and challenged with ELISA, qPCR, or culture methods. This work enriches the arsenal of signal transduction based on aptamer epitope recognition, highlighting its potential as a precision on-site diagnostic tool for pathogenic bacteria.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"1 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144566164","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}
Yang Li, Yixuan Bian, Regan McMath, James Davis, Pagona Papakonstantinou, Yuanhua Shao, Meixian Li
{"title":"Construction of Flexible Electrochemical Sensing Interfaces Based on Paper/PEDOT:PSS for the Detection of Cystatin C.","authors":"Yang Li, Yixuan Bian, Regan McMath, James Davis, Pagona Papakonstantinou, Yuanhua Shao, Meixian Li","doi":"10.1021/acs.analchem.5c01627","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01627","url":null,"abstract":"<p><p>Chronic kidney disease (CKD) is a global public health issue, with high prevalence and mortality. As cystatin C is a biomarker for the early diagnosis of CKD, there is an urgent need to develop a simple and inexpensive method for the detection of Cys C in human plasma. Herein, a sensitive method with flexible electrochemical aptamer-based (E-AB) sensing interfaces has been established using poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate)-treated paper-based electrodes. Introduction of MoS<sub>2</sub> quantum dots increased the specific surface area and realized effective immobilization of the thiolated Cys C aptamer (Cys C Apt) modified with methylene blue through the formation of disulfide bonds. The E-AB sensing interfaces were successfully applied to the quantification of Cys C in human venous whole blood and plasma samples based on the binding-induced conformational change mechanism of Cys C Apt by employing square wave voltammetry. The flexible E-AB sensing interfaces show significant potential to be developed into wearable patches for in vivo detection of Cys C, which will pave the way for early diagnosis and long-term monitoring of CKD.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":" ","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558400","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}
Qi Zhu, Xu Sun, Yu Du, Xiaoyue Zhang, Xiang Ren, Hongmin Ma, Dan Wu, Huangxian Ju, Qin Wei
{"title":"Detection of Matrix Metalloproteinase 2 by a Polypeptide Cleavable Split-Type Sensor Based on Atomically Precise Bimetallic Nanocluster Au<sub>3</sub>Ag<sub>5</sub>(MSA)<sub>3</sub>.","authors":"Qi Zhu, Xu Sun, Yu Du, Xiaoyue Zhang, Xiang Ren, Hongmin Ma, Dan Wu, Huangxian Ju, Qin Wei","doi":"10.1021/acs.analchem.5c02007","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c02007","url":null,"abstract":"<p><p>In this study, a split-type electrochemiluminescence (ECL) sensor based on polypeptide cleavage was proposed for the sensitive detection of matrix metalloproteinase 2 (MMP 2). The bimetallic nanoclusters were protected by mercaptosuccinic acid (MSA), named Au<sub>3</sub>Ag<sub>5</sub>(MSA)<sub>3</sub>, serving as a signal probe. Owing to the synergistic effect of the bimetal composition, the clusters exhibited enhanced ECL properties. Polypeptide chains (PLGVR) that can be specifically cleaved by MMP 2 were selected from the MMP 2 domain. Additionally, the antifouling segment (PPEKEK) and the binding segment (CCC) were incorporated into the polypeptide chain design. In the presence of MMP 2, a specific cleavage site on the polypeptide was targeted, resulting in the detachment of Au<sub>3</sub>Ag<sub>5</sub>(MSA)<sub>3</sub> and subsequent generation of an ECL signal. Compared to conventional detection methods, this approach exhibited enhanced sensitivity and improved stability. This method provides a valuable reference for detecting members of the matrix metalloproteinase family and holds promising prospects for future development.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":" ","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564155","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}