Glycoproteomics has emerged as a powerful tool for decoding cellular communication and disease mechanisms. However, the structural complexity and low abundance of glycoproteins demand high-efficiency enrichment materials. Conventional methods such as lectin affinity, hydrazide chemistry, metal ion affinity, and hydrophilic interaction chromatography have been widely used, but they suffer from drawbacks including limited specificity, harsh chemical conditions, or nonspecific adsorption. These constraints restrict their utility for complex biological specimens and impede clinical translation. Boronate affinity materials, which enable reversible and selective binding to cis-diol moieties, present a promising alternative for glycoprotein isolation.
Results
Recent advances in boronate-affinity materials have led to notable improvements in specificity, stability, and biocompatibility, substantially broadening their applicability in glycoproteomics. Mesoporous frameworks provide high surface areas and tunable pores that enhance adsorption capacity and facilitate efficient capture of glycoproteins. Boronate-functionalized nanocomposites exhibit superior responsiveness to physiological pH and temperature, ensuring stable performance in complex biological environments. Molecularly imprinted polymers, with their tailor-made recognition sites, achieve exceptional selectivity toward glycoprotein motifs. Together, these innovations enable high-throughput, sensitive, and reproducible enrichment of glycoproteins, even from challenging samples such as plasma, cerebrospinal fluid, and exosomes. Importantly, enrichment via these strategies has facilitated the identification of clinically relevant glycoprotein biomarkers associated with cancer, neurodegenerative disorders, and metabolic diseases. Moreover, the integration of boronate affinity enrichment with advanced analytical techniques, such as mass spectrometry and microfluidics, further underscores its potential to improve diagnostic accuracy and accelerate clinical translation in precision medicine.
Significance
This review highlights the unique advantages of boronate affinity materials in addressing the limitations of conventional glycoprotein enrichment. Their efficiency, specificity, and clinical adaptability not only facilitate biomarker discovery but also promote the integration of glycoproteomics into next-generation precision diagnostics. In addition, by summarizing the design principles, functional mechanisms, and biomedical applications of these materials, this review provides a framework to support their further development and clinical translation.
{"title":"Recent advances and clinical perspectives of boronate affinity-driven glycoproteomics: A review","authors":"Huiying Zhang, Yufei Shi, Fenghua Xu, Xin Dong, Lijun Sun, Xue Zhang","doi":"10.1016/j.aca.2025.344976","DOIUrl":"https://doi.org/10.1016/j.aca.2025.344976","url":null,"abstract":"<h3>Background</h3>Glycoproteomics has emerged as a powerful tool for decoding cellular communication and disease mechanisms. However, the structural complexity and low abundance of glycoproteins demand high-efficiency enrichment materials. Conventional methods such as lectin affinity, hydrazide chemistry, metal ion affinity, and hydrophilic interaction chromatography have been widely used, but they suffer from drawbacks including limited specificity, harsh chemical conditions, or nonspecific adsorption. These constraints restrict their utility for complex biological specimens and impede clinical translation. Boronate affinity materials, which enable reversible and selective binding to cis-diol moieties, present a promising alternative for glycoprotein isolation.<h3>Results</h3>Recent advances in boronate-affinity materials have led to notable improvements in specificity, stability, and biocompatibility, substantially broadening their applicability in glycoproteomics. Mesoporous frameworks provide high surface areas and tunable pores that enhance adsorption capacity and facilitate efficient capture of glycoproteins. Boronate-functionalized nanocomposites exhibit superior responsiveness to physiological pH and temperature, ensuring stable performance in complex biological environments. Molecularly imprinted polymers, with their tailor-made recognition sites, achieve exceptional selectivity toward glycoprotein motifs. Together, these innovations enable high-throughput, sensitive, and reproducible enrichment of glycoproteins, even from challenging samples such as plasma, cerebrospinal fluid, and exosomes. Importantly, enrichment via these strategies has facilitated the identification of clinically relevant glycoprotein biomarkers associated with cancer, neurodegenerative disorders, and metabolic diseases. Moreover, the integration of boronate affinity enrichment with advanced analytical techniques, such as mass spectrometry and microfluidics, further underscores its potential to improve diagnostic accuracy and accelerate clinical translation in precision medicine.<h3>Significance</h3>This review highlights the unique advantages of boronate affinity materials in addressing the limitations of conventional glycoprotein enrichment. Their efficiency, specificity, and clinical adaptability not only facilitate biomarker discovery but also promote the integration of glycoproteomics into next-generation precision diagnostics. In addition, by summarizing the design principles, functional mechanisms, and biomedical applications of these materials, this review provides a framework to support their further development and clinical translation.","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"223 1","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145664849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nitrogenous bases, namely purine and pyrimidine, and their derivatives are key metabolites for the growth and division of cells as they are an integral part of biomolecules that store genetic information, involved in protein synthesis, as energy carrier molecules, and many other metabolic processes. Purine and pyrimidine can either be synthesized (de novo pathway), recycled, and/or acquired exogenously (salvage pathway). These nitrogenous bases can be analysed with high sensitivity using mass spectrometry-based methods, which are inherently destructive. Therefore, there remains a challenge for imaging of nascent purine and pyrimidine nitrogenous bases distribution at the single-cell level.
Results
Here, we report a novel single-cell Raman imaging framework, “carbon isotope imaging and spectral tracing (CIIST)” for mapping nascent nitrogenous bases in prokaryotic and eukaryotic microbial systems. This method helps in visualising the turnover of nascent purine and pyrimidine nitrogenous bases at the subcellular level over time. The enrichment of carbon isotope (carbon-12 or carbon-13) in the nitrogenous base pool generates Raman peaks at distinct positions, and targeting these CIIST can be used for generating spatial maps for quasi-quantitative imaging of nitrogenous base turnover in cells.
Significance
Overall findings provide the prospective utility of the CIIST technique as a highly effective non-destructive tool for multiplex and spatial biomolecular analysis of nitrogenous base metabolism at the single-cell level.
{"title":"Visualization of de novo synthesized nascent purine and pyrimidine using carbon isotope imaging and spectral tracing (CIIST) in single microbial cell","authors":"Jiro Karlo, Tejas Ajit Mhaiskar, Hrishikesh Ravindra Karande, S.P. Singh","doi":"10.1016/j.aca.2025.344986","DOIUrl":"10.1016/j.aca.2025.344986","url":null,"abstract":"<div><h3>Background</h3><div>Nitrogenous bases, namely purine and pyrimidine, and their derivatives are key metabolites for the growth and division of cells as they are an integral part of biomolecules that store genetic information, involved in protein synthesis, as energy carrier molecules, and many other metabolic processes. Purine and pyrimidine can either be synthesized (<em>de novo</em> pathway), recycled, and/or acquired exogenously (salvage pathway). These nitrogenous bases can be analysed with high sensitivity using mass spectrometry-based methods, which are inherently destructive. Therefore, there remains a challenge for imaging of nascent purine and pyrimidine nitrogenous bases distribution at the single-cell level.</div></div><div><h3>Results</h3><div>Here, we report a novel single-cell Raman imaging framework, “carbon isotope imaging and spectral tracing (CIIST)” for mapping nascent nitrogenous bases in prokaryotic and eukaryotic microbial systems. This method helps in visualising the turnover of nascent purine and pyrimidine nitrogenous bases at the subcellular level over time. The enrichment of carbon isotope (carbon-12 or carbon-13) in the nitrogenous base pool generates Raman peaks at distinct positions, and targeting these CIIST can be used for generating spatial maps for quasi-quantitative imaging of nitrogenous base turnover in cells.</div></div><div><h3>Significance</h3><div>Overall findings provide the prospective utility of the CIIST technique as a highly effective non-destructive tool for multiplex and spatial biomolecular analysis of nitrogenous base metabolism at the single-cell level.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344986"},"PeriodicalIF":6.0,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145690027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-03DOI: 10.1016/j.aca.2025.344984
Kai Wang , Dan-Xian Cai , Hao-Xiang Jiang , Zi-Rui Luo , Hong-Rui Zhan
Sulfite (SO32−) plays a crucial role in the transfer of substances and information between organisms and their environments. This study developed a novel hemicyanine-based sensor, named HD-TT, for the real-time and highly sensitive detection of to track the SO32− in the biosystem. The HD-TT probe is selectively activated by SO32−, resulting in simultaneous enhancement of both absorption and fluorescence signals. It displays a linear response to sulfite concentrations and achieves a detection limit of 151.61 nM, reflecting high sensitivity under physiologically relevant conditions. In cellular models, HD-TT effectively monitored dynamic changes in SO32− levels, confirming its capability for real-time tracking in biological environments. In addition, the probe successfully expressed a remarked fluorescence intensity in RA serum samples, underscoring its detecting potential towards SO32−. Furthermore, the probe also exhibited an excellent specificity against common biological interferents and maintained stability across a range of physiological conditions, supporting its suitability in biosystem. These attributes position HD-TT as a promising non-invasive tool for dynamic monitoring SO32−, facilitating an novel perspective in biomedical research.
{"title":"A near-infrared hemicyanine-derived fluorescent probe for sulfite detection in biomedical investigate","authors":"Kai Wang , Dan-Xian Cai , Hao-Xiang Jiang , Zi-Rui Luo , Hong-Rui Zhan","doi":"10.1016/j.aca.2025.344984","DOIUrl":"10.1016/j.aca.2025.344984","url":null,"abstract":"<div><div>Sulfite (SO<sub>3</sub><sup>2−</sup>) plays a crucial role in the transfer of substances and information between organisms and their environments. This study developed a novel hemicyanine-based sensor, named <strong>HD-TT</strong>, for the real-time and highly sensitive detection of to track the SO<sub>3</sub><sup>2−</sup> in the biosystem. The <strong>HD-TT</strong> probe is selectively activated by SO<sub>3</sub><sup>2−</sup>, resulting in simultaneous enhancement of both absorption and fluorescence signals. It displays a linear response to sulfite concentrations and achieves a detection limit of 151.61 nM, reflecting high sensitivity under physiologically relevant conditions. In cellular models, <strong>HD-TT</strong> effectively monitored dynamic changes in SO<sub>3</sub><sup>2−</sup> levels, confirming its capability for real-time tracking in biological environments. In addition, the probe successfully expressed a remarked fluorescence intensity in RA serum samples, underscoring its detecting potential towards SO<sub>3</sub><sup>2−</sup>. Furthermore, the probe also exhibited an excellent specificity against common biological interferents and maintained stability across a range of physiological conditions, supporting its suitability in biosystem. These attributes position <strong>HD-TT</strong> as a promising non-invasive tool for dynamic monitoring SO<sub>3</sub><sup>2−</sup>, facilitating an novel perspective in biomedical research.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344984"},"PeriodicalIF":6.0,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145674119","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-03DOI: 10.1016/j.aca.2025.344952
Zuzana Lásko , Ondřej Peterka , Robert Jirásko , Anna Taylor , Tomáš Hájek , Beatrice Mohelníková-Duchoňová , Martin Loveček , Bohuslav Melichar , Michal Holčapek
Background
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers, mainly due to the late diagnosis and the lack of reliable biomarkers. Lipidomics provides a promising approach for identifying disease-related alterations, but existing methods are often limited to lipid class profiles with insufficient molecular detail. Reversed-phase ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry (RP-UHPLC/MS/MS) offers the possibility to determine lipids at the fatty acyl/alkyl level. Here, we address the need for a validated quantitative workflow that enables accurate and reproducible lipidomic profiling of human serum in the context of PDAC.
Results
We developed and validated an RP-UHPLC/MS/MS method using multiple reaction monitoring, enabling the identification of 455 lipid species from 22 subclasses, with 381 species from 21 subclasses quantified. The workflow included a response factor correction for sterol esters, which markedly improved their quantification accuracy. The application to serum samples from 54 PDAC patients and 55 healthy controls yielded highly reproducible data, with clear group separation observed in both unsupervised and supervised statistical analyses. Dysregulation was most prominent in sphingolipids and phospholipids. Very long-chain saturated sphingolipids (≥C22) were downregulated, while some shorter or unsaturated chains showed mild upregulation. Phospholipid alterations were dominated by species containing polyunsaturated fatty acyls, particularly 18:2 and 20:4, with plasmalogens showing the strongest changes. These structurally resolved findings were further supported by gas chromatography – mass spectrometry analysis of fatty acid methyl esters.
Significance
This validated workflow provides comprehensive quantitative coverage across 21 lipid subclasses with the structural resolution critical for biological interpretation. The detailed mapping of sphingolipid and phospholipid dysregulation in PDAC demonstrates that only the fatty acyl level annotation reveals molecular signatures that may reflect specific enzymatic activities or pathways. The method delivers a robust platform for biomarker discovery and mechanistic studies in cancer lipidomics.
{"title":"RP-UHPLC/MS/MS provides enhanced lipidomic profiling of human serum in pancreatic cancer","authors":"Zuzana Lásko , Ondřej Peterka , Robert Jirásko , Anna Taylor , Tomáš Hájek , Beatrice Mohelníková-Duchoňová , Martin Loveček , Bohuslav Melichar , Michal Holčapek","doi":"10.1016/j.aca.2025.344952","DOIUrl":"10.1016/j.aca.2025.344952","url":null,"abstract":"<div><h3>Background</h3><div>Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers, mainly due to the late diagnosis and the lack of reliable biomarkers. Lipidomics provides a promising approach for identifying disease-related alterations, but existing methods are often limited to lipid class profiles with insufficient molecular detail. Reversed-phase ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry (RP-UHPLC/MS/MS) offers the possibility to determine lipids at the fatty acyl/alkyl level. Here, we address the need for a validated quantitative workflow that enables accurate and reproducible lipidomic profiling of human serum in the context of PDAC.</div></div><div><h3>Results</h3><div>We developed and validated an RP-UHPLC/MS/MS method using multiple reaction monitoring, enabling the identification of 455 lipid species from 22 subclasses, with 381 species from 21 subclasses quantified. The workflow included a response factor correction for sterol esters, which markedly improved their quantification accuracy. The application to serum samples from 54 PDAC patients and 55 healthy controls yielded highly reproducible data, with clear group separation observed in both unsupervised and supervised statistical analyses. Dysregulation was most prominent in sphingolipids and phospholipids. Very long-chain saturated sphingolipids (≥C22) were downregulated, while some shorter or unsaturated chains showed mild upregulation. Phospholipid alterations were dominated by species containing polyunsaturated fatty acyls, particularly 18:2 and 20:4, with plasmalogens showing the strongest changes. These structurally resolved findings were further supported by gas chromatography – mass spectrometry analysis of fatty acid methyl esters.</div></div><div><h3>Significance</h3><div>This validated workflow provides comprehensive quantitative coverage across 21 lipid subclasses with the structural resolution critical for biological interpretation. The detailed mapping of sphingolipid and phospholipid dysregulation in PDAC demonstrates that only the fatty acyl level annotation reveals molecular signatures that may reflect specific enzymatic activities or pathways. The method delivers a robust platform for biomarker discovery and mechanistic studies in cancer lipidomics.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344952"},"PeriodicalIF":6.0,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145657593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-02DOI: 10.1016/j.aca.2025.344983
Xiaoshuang Chen , Junxia Han , Mingyuan Sun , Zhenhua Wang , Shuai Wang , Lin Han , Yu Zhang
Background
Extracellular vesicles (EVs) serve as crucial biomarkers for cancer screening due to their close association with the physiological and pathological states of cancer cells. These membrane-bound vesicles carry molecular cargo that reflects the characteristics of their parent cells, making them valuable diagnostic indicators. Current EVs detection methods face significant limitations in clinical applications, particularly regarding sensitivity and accuracy requirements for reliable cancer diagnostics. The critical challenge is to develop sensitive and accurate EVs detection methods for clinical cancer screening.
Results
We developed an aptamer-integrated dual-mode microfluidic biosensor combining electrochemistry (EC) and photoluminescence (PL) detection for ovarian cancer EVs screening. The platform utilized aptamer-functionalized gold nanoflowers (Au NFs) integrated with 3D laser-induced graphene (LIG) electrode arrays, achieving a 1.44-fold increase in electroactive surface area. Poly-lysine (PLL) served as the PL detection substrate for capturing fluorescent complexes, enabling simultaneous dual-signal generation. Under optimized conditions, the biosensor achieved a detection range of 10-106 particles/μL with a detection limit of 33 particles/μL. Clinical validation demonstrated excellent accuracy with spiked recoveries of 98.6 %–101.8 % and successful application in ovarian cancer patient samples, confirming practical diagnostic utility.
Significance
This work presents the first aptamer-integrated EC-PL dual-mode microfluidic biosensor for EVs detection, enabling real-time signal cross-validation and enhanced reliability. The novel combination of Au NFs-3D LIG architecture with PLL-mediated detection strategy significantly improves sensitivity and clinical applicability for point-of-care cancer screening.
外胞囊泡（EVs）与癌细胞的生理和病理状态密切相关，是癌症筛查的重要生物标志物。这些膜结合的囊泡携带反映其亲本细胞特征的分子货物，使其成为有价值的诊断指标。目前的EV检测方法在临床应用中面临很大的局限性，特别是在可靠的癌症诊断的敏感性和准确性要求方面。关键的挑战是开发敏感和准确的ev检测方法用于临床癌症筛查。结果建立了一种结合电化学（EC）和光致发光（PL）检测的适配体集成双模微流控生物传感器，用于卵巢癌EV筛查。该平台将适配体功能化的金纳米花（Au NFs）与3D激光诱导石墨烯（LIG）电极阵列集成在一起，使电活性表面积增加了1.44倍。聚赖氨酸（PLL）作为PL检测底物捕获荧光复合物，实现双信号同时产生。在优化条件下，该传感器的检测范围为10 ~ 106个粒子/μL，检出限为33个粒子/μL。临床验证结果表明，加标回收率为98.6% ~ 101.8%，具有良好的准确性，在卵巢癌患者样品中得到了成功的应用，具有实际的诊断价值。本工作提出了首个用于电动汽车检测的适配体集成EC-PL双模微流控生物传感器，实现了实时信号交叉验证，提高了可靠性。Au nf - 3d LIG结构与pll介导的检测策略的新组合显着提高了即时癌症筛查的敏感性和临床适用性。
{"title":"An aptamer integrated electrochemistry and photoluminescence dual-mode microfluidic biosensor for sensitive and accurate ovarian cancer extracellular vesicles detection","authors":"Xiaoshuang Chen , Junxia Han , Mingyuan Sun , Zhenhua Wang , Shuai Wang , Lin Han , Yu Zhang","doi":"10.1016/j.aca.2025.344983","DOIUrl":"10.1016/j.aca.2025.344983","url":null,"abstract":"<div><h3>Background</h3><div>Extracellular vesicles (EVs) serve as crucial biomarkers for cancer screening due to their close association with the physiological and pathological states of cancer cells. These membrane-bound vesicles carry molecular cargo that reflects the characteristics of their parent cells, making them valuable diagnostic indicators. Current EVs detection methods face significant limitations in clinical applications, particularly regarding sensitivity and accuracy requirements for reliable cancer diagnostics. The critical challenge is to develop sensitive and accurate EVs detection methods for clinical cancer screening.</div></div><div><h3>Results</h3><div>We developed an aptamer-integrated dual-mode microfluidic biosensor combining electrochemistry (EC) and photoluminescence (PL) detection for ovarian cancer EVs screening. The platform utilized aptamer-functionalized gold nanoflowers (Au NFs) integrated with 3D laser-induced graphene (LIG) electrode arrays, achieving a 1.44-fold increase in electroactive surface area. Poly-lysine (PLL) served as the PL detection substrate for capturing fluorescent complexes, enabling simultaneous dual-signal generation. Under optimized conditions, the biosensor achieved a detection range of 10-10<sup>6</sup> particles/μL with a detection limit of 33 particles/μL. Clinical validation demonstrated excellent accuracy with spiked recoveries of 98.6 %–101.8 % and successful application in ovarian cancer patient samples, confirming practical diagnostic utility.</div></div><div><h3>Significance</h3><div>This work presents the first aptamer-integrated EC-PL dual-mode microfluidic biosensor for EVs detection, enabling real-time signal cross-validation and enhanced reliability. The novel combination of Au NFs-3D LIG architecture with PLL-mediated detection strategy significantly improves sensitivity and clinical applicability for point-of-care cancer screening.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344983"},"PeriodicalIF":6.0,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-02DOI: 10.1016/j.aca.2025.344979
Yuan Kong , Zhaoying Liu , Guizhu Wu , Xiaoyong Liu , Song Fang , Kun Zeng , Zhen Zhang , Ming Li
Background
Portable biosensing strategies are promising rapid and on-site detection methods, possessing the efficiency and accessibility for detecting target. Hydrogel present numerous advantages in rapid analysis owing to their remarkable biological loading capabilities and optical properties. However, traditional hydrogels suffer from some drawbacks, such as poor signal intensity and slow responsiveness, restricting their further utilization in practice.
Results
To address these challenges, an enhanced DNA cryogel biosensing platform based on dual-RCA and dual-enzyme had proposed for quantification of zearalenone (ZEN) in this study. The MB@Apt-cDNA probe and DNA cryogel were prepared in advance to provide accurate recognition and portable interface. When ZEN presented, the released cDNA aroused rolling-circle amplification (RCA) to amplify signal, and then introduced dual-enzyme system by coupling glucose oxidase and horseradish peroxidase. On the dual-RCA-based DNA cryogel, the coloration in situ displayed under self-powered H2O2 system. The results identified through visualization, smartphone and color chart, exhibiting a limit of detection (LOD) at 6.95 pg/mL, with detection range between 10 pg/mL to 20 ng/mL. This biosensing showed satisfactory specificity, accuracy, efficiency and accessibility by its application in spiked and real samples. Furthermore, the highlights, including high integration, simplified operation and portable reading, had demonstrated.
Significance
The enhanced chromogenic cryogel platform can provide by integrating advanced strategies, which may have promising prospects for portable monitoring.
{"title":"Enhanced DNA cryogel platform: Dual-RCA-Driven coloration in situ for portable biosensing","authors":"Yuan Kong , Zhaoying Liu , Guizhu Wu , Xiaoyong Liu , Song Fang , Kun Zeng , Zhen Zhang , Ming Li","doi":"10.1016/j.aca.2025.344979","DOIUrl":"10.1016/j.aca.2025.344979","url":null,"abstract":"<div><h3>Background</h3><div>Portable biosensing strategies are promising rapid and on-site detection methods, possessing the efficiency and accessibility for detecting target. Hydrogel present numerous advantages in rapid analysis owing to their remarkable biological loading capabilities and optical properties. However, traditional hydrogels suffer from some drawbacks, such as poor signal intensity and slow responsiveness, restricting their further utilization in practice.</div></div><div><h3>Results</h3><div>To address these challenges, an enhanced DNA cryogel biosensing platform based on dual-RCA and dual-enzyme had proposed for quantification of zearalenone (ZEN) in this study. The MB@Apt-cDNA probe and DNA cryogel were prepared in advance to provide accurate recognition and portable interface. When ZEN presented, the released cDNA aroused rolling-circle amplification (RCA) to amplify signal, and then introduced dual-enzyme system by coupling glucose oxidase and horseradish peroxidase. On the dual-RCA-based DNA cryogel, the coloration in situ displayed under self-powered H<sub>2</sub>O<sub>2</sub> system. The results identified through visualization, smartphone and color chart, exhibiting a limit of detection (LOD) at 6.95 pg/mL, with detection range between 10 pg/mL to 20 ng/mL. This biosensing showed satisfactory specificity, accuracy, efficiency and accessibility by its application in spiked and real samples. Furthermore, the highlights, including high integration, simplified operation and portable reading, had demonstrated.</div></div><div><h3>Significance</h3><div>The enhanced chromogenic cryogel platform can provide by integrating advanced strategies, which may have promising prospects for portable monitoring.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344979"},"PeriodicalIF":6.0,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-02DOI: 10.1016/j.aca.2025.344980
Chanyong Park , Dongju Ha , Seoyeon Bae , Soo-Hwan Jeong , Sangmin Jeon , Yoochan Hong , Ohwon Kwon , Dongkyu Lee
Background
Rapid and accurate determination of bacterial antibiotic susceptibility is essential for effective therapy and prevention of antimicrobial resistance. However, conventional culture-based assays are time-consuming (≥24 h) and labor-intensive, requiring centralized infrastructure. While rapid approaches such as microfluidics and mass spectrometry have been developed, they are often limited by high cost, complex instrumentation, or poor portability, underscoring the need for simple, field-deployable alternatives.
Results
We developed a miniaturized platform integrating a volatile organic compound (VOC) sensor with thermal regulation and onboard agitation to monitor bacterial growth and antibiotic susceptibility in real time. Optimization of glucose concentration and agitation allowed E. coli O157:H7 to be incubated in sealed vials, with VOC emissions recorded every second. Lag-phase analysis of VOC curves, modeled using a modified Gompertz function, quantitatively estimated bacterial concentration. Antibiotic exposure caused dose-dependent delays or complete suppression of VOC signals, producing MICs of ∼1 μg/mL for enrofloxacin, 8 μg/mL for gentamicin, and 0.7 μg/mL for cefoperazone. Susceptible and resistant phenotypes were correctly classified within 12 h. The platform maintained performance in opaque matrices such as milk without preprocessing and was validated with non-pathogenic E. coli and Salmonella typhimurium, demonstrating accurate detection and strain-specific susceptibility profiles.
Significance
This VOC-based miniaturized AST platform enables rapid, label-free assessment of bacterial growth and susceptibility in under 12 h without optics, plating, or extensive sample processing. Its portability and compatibility with diverse pathogens, antibiotics, and opaque samples position it as a promising tool for decentralized diagnostics, point-of-care testing, and food-safety monitoring.
{"title":"Miniaturized volatile organic compound sensor platform for the real-time monitoring of bacterial growth and antibiotic susceptibility","authors":"Chanyong Park , Dongju Ha , Seoyeon Bae , Soo-Hwan Jeong , Sangmin Jeon , Yoochan Hong , Ohwon Kwon , Dongkyu Lee","doi":"10.1016/j.aca.2025.344980","DOIUrl":"10.1016/j.aca.2025.344980","url":null,"abstract":"<div><h3>Background</h3><div>Rapid and accurate determination of bacterial antibiotic susceptibility is essential for effective therapy and prevention of antimicrobial resistance. However, conventional culture-based assays are time-consuming (≥24 h) and labor-intensive, requiring centralized infrastructure. While rapid approaches such as microfluidics and mass spectrometry have been developed, they are often limited by high cost, complex instrumentation, or poor portability, underscoring the need for simple, field-deployable alternatives.</div></div><div><h3>Results</h3><div>We developed a miniaturized platform integrating a volatile organic compound (VOC) sensor with thermal regulation and onboard agitation to monitor bacterial growth and antibiotic susceptibility in real time. Optimization of glucose concentration and agitation allowed <em>E. coli</em> O157:H7 to be incubated in sealed vials, with VOC emissions recorded every second. Lag-phase analysis of VOC curves, modeled using a modified Gompertz function, quantitatively estimated bacterial concentration. Antibiotic exposure caused dose-dependent delays or complete suppression of VOC signals, producing MICs of ∼1 μg/mL for enrofloxacin, 8 μg/mL for gentamicin, and 0.7 μg/mL for cefoperazone. Susceptible and resistant phenotypes were correctly classified within 12 h. The platform maintained performance in opaque matrices such as milk without preprocessing and was validated with non-pathogenic <em>E. coli</em> and <em>Salmonella typhimurium</em>, demonstrating accurate detection and strain-specific susceptibility profiles.</div></div><div><h3>Significance</h3><div>This VOC-based miniaturized AST platform enables rapid, label-free assessment of bacterial growth and susceptibility in under 12 h without optics, plating, or extensive sample processing. Its portability and compatibility with diverse pathogens, antibiotics, and opaque samples position it as a promising tool for decentralized diagnostics, point-of-care testing, and food-safety monitoring.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1385 ","pages":"Article 344980"},"PeriodicalIF":6.0,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145657598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-02DOI: 10.1016/j.aca.2025.344981
Wenchao Geng , Jiangying Ji , Zhiyi Yan , Fan Wu , Jinyuan Chang , Jiarui Wei , Mengjiao Mei , Shang Chen , Fei Wang , Ruiying Yang
Background
Exosomal long non-coding RNA (lncRNA) as a burgeoning biomarker, its abnormal expression is closely related to the progression of cancer. Therefore, the development of sensitive exosomal lncRNA detection methods has significant effect in promoting the early diagnosis of cancers. Herein, a novel signal-on photoelectrochemical (PEC) biosensor is reported for the highly sensitive detection of exosomal lncRNA combined porous COF/TiO2 nanospheres and terminal deoxynucleotidyl transferase (TdT)-triggered poly(C) with silver nanoclusters (DNA-AgCNs) nanowires, using machine learning to assist in cancer intelligent diagnosis.
Results
The as-prepared COF/TiO2 nanospheres as photoelectrode materials with high specific surface area and porosity can carry abundant tyrosine (Tyr). Then, target lncRNA HOTAIR can bind with phosphate groups at the 3′-end of DNA (pDNA) to expose the porous sites to adsorb Tyr. Subsequently, TdT can catalyze deoxycytidine triphosphate (dCTP) cyclic amplification to in-situ generate abundant DNA-AgCNs nanowires, which can act as a bridge for electron transfer, enhancing the photocurrent of COF/TiO2 nanospheres. The developed PEC biosensor achieves a wide range from 500 aM to 10 pM with a low detection limit of 116 aM. Importantly, this machine learning is employed to probe the hidden potential pattern in the developed PEC data, and machine learning for cancer intelligent diagnosis can achieve 85.7 % accuracy, 100 % sensitivity and 80.0 % specificity.
Significance
Machine learning-assisted PEC biosensor based on DNA-AgNCs nanowires and porous COF/TiO2 nanospheres can effectively distinguish the expressions of lncRNA HOTAIR in plasma exosomes from healthy people and cancer patients, which not only significantly enhances the sensitivity and accuracy of cancer diagnosis, but also provides a great application prospect in the early diagnosis of lncRNA-related cancer.
{"title":"Machine learning-assisted photoelectrochemical biosensor based on DNA-AgNCs nanowires for exosomal lncRNA intelligent diagnosis","authors":"Wenchao Geng , Jiangying Ji , Zhiyi Yan , Fan Wu , Jinyuan Chang , Jiarui Wei , Mengjiao Mei , Shang Chen , Fei Wang , Ruiying Yang","doi":"10.1016/j.aca.2025.344981","DOIUrl":"10.1016/j.aca.2025.344981","url":null,"abstract":"<div><h3>Background</h3><div>Exosomal long non-coding RNA (lncRNA) as a burgeoning biomarker, its abnormal expression is closely related to the progression of cancer. Therefore, the development of sensitive exosomal lncRNA detection methods has significant effect in promoting the early diagnosis of cancers. Herein, a novel signal-on photoelectrochemical (PEC) biosensor is reported for the highly sensitive detection of exosomal lncRNA combined porous COF/TiO<sub>2</sub> nanospheres and terminal deoxynucleotidyl transferase (TdT)-triggered poly(C) with silver nanoclusters (DNA-AgCNs) nanowires, using machine learning to assist in cancer intelligent diagnosis.</div></div><div><h3>Results</h3><div>The as-prepared COF/TiO<sub>2</sub> nanospheres as photoelectrode materials with high specific surface area and porosity can carry abundant tyrosine (Tyr). Then, target lncRNA HOTAIR can bind with phosphate groups at the 3′-end of DNA (pDNA) to expose the porous sites to adsorb Tyr. Subsequently, TdT can catalyze deoxycytidine triphosphate (dCTP) cyclic amplification to in-situ generate abundant DNA-AgCNs nanowires, which can act as a bridge for electron transfer, enhancing the photocurrent of COF/TiO<sub>2</sub> nanospheres. The developed PEC biosensor achieves a wide range from 500 aM to 10 pM with a low detection limit of 116 aM. Importantly, this machine learning is employed to probe the hidden potential pattern in the developed PEC data, and machine learning for cancer intelligent diagnosis can achieve 85.7 % accuracy, 100 % sensitivity and 80.0 % specificity.</div></div><div><h3>Significance</h3><div>Machine learning-assisted PEC biosensor based on DNA-AgNCs nanowires and porous COF/TiO<sub>2</sub> nanospheres can effectively distinguish the expressions of lncRNA HOTAIR in plasma exosomes from healthy people and cancer patients, which not only significantly enhances the sensitivity and accuracy of cancer diagnosis, but also provides a great application prospect in the early diagnosis of lncRNA-related cancer.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344981"},"PeriodicalIF":6.0,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01DOI: 10.1016/j.aca.2025.344967
Zixi Li , Zhicheng Huang , Daoyun Wang , Tong Li , Junya Peng , Anlan Zhang , Ruimeng Sun , Xin Wu , Yu Zhang , Naixin Liang , Qin Li , Zewen Wei
Background
The beating of motile cilia plays an important role in many physiological processes. In recent years, the precise monitoring of ciliary beating has provided valuable insights into cilia-related diseases and their therapies. However, a gap persists between homogeneous in vitro cell models and heterogeneous in vivo ciliary systems. Human-derived ciliary organoids (HDCOs) show promise in bridging this gap but face technical barriers: limited numbers of HDCOs cannot meet multi-well-plate demands, and long-term tracking accuracy is compromised by organoid overlapping/displacement during culture.
Results
A multifunctional microfluidic chip (MOCiB-Chip) was developed, featuring a three-layer structure (fluidic layer, porous membrane, capture layer) and two independent regions for multi-concentration experiments. It integrates single-HDCO immobilization, culture, in situ observation, and on-chip drug mixing. Combined with an independently developed analysis program, Ciliary Beating Frequency Analysis Software (CBFAS), and a fluid control module, the automatic quantifying system for ciliary beating (AuCilia) was established. It achieved 7-day monitoring of human bronchial HDCOs and evaluated roflumilast at 0–100 nM: roflumilast at 10 nM increased beating frequency by 81.5 %, 1 nM by 27.8 %, while 100 nM caused cessation by day 2. cAMP measurements confirmed roflumilast's mechanism, validating the system.
Significance
This strategy eliminates HDCO loss via on-chip integration of all procedures, resolving the conflict between limited samples and multi-condition tests. It improves tracking accuracy by avoiding organoid displacement/overlapping, overcoming multi-well-plate limitations. The system's low cost and high reproducibility accelerate drug screening (e.g., roflumilast) and enable precise study of cilia physiology, laying a key technical foundation for cilia-related disease research and clinical translation.
{"title":"Organoid chip based automatic system for long-term quantifying ciliary beating under drug intervention","authors":"Zixi Li , Zhicheng Huang , Daoyun Wang , Tong Li , Junya Peng , Anlan Zhang , Ruimeng Sun , Xin Wu , Yu Zhang , Naixin Liang , Qin Li , Zewen Wei","doi":"10.1016/j.aca.2025.344967","DOIUrl":"10.1016/j.aca.2025.344967","url":null,"abstract":"<div><h3>Background</h3><div>The beating of motile cilia plays an important role in many physiological processes. In recent years, the precise monitoring of ciliary beating has provided valuable insights into cilia-related diseases and their therapies. However, a gap persists between homogeneous <em>in vitro</em> cell models and heterogeneous <em>in vivo</em> ciliary systems. Human-derived ciliary organoids (HDCOs) show promise in bridging this gap but face technical barriers: limited numbers of HDCOs cannot meet multi-well-plate demands, and long-term tracking accuracy is compromised by organoid overlapping/displacement during culture.</div></div><div><h3>Results</h3><div>A multifunctional microfluidic chip (MOCiB-Chip) was developed, featuring a three-layer structure (fluidic layer, porous membrane, capture layer) and two independent regions for multi-concentration experiments. It integrates single-HDCO immobilization, culture, <em>in situ</em> observation, and on-chip drug mixing. Combined with an independently developed analysis program, Ciliary Beating Frequency Analysis Software (CBFAS), and a fluid control module, the automatic quantifying system for ciliary beating (AuCilia) was established. It achieved 7-day monitoring of human bronchial HDCOs and evaluated roflumilast at 0–100 nM: roflumilast at 10 nM increased beating frequency by 81.5 %, 1 nM by 27.8 %, while 100 nM caused cessation by day 2. cAMP measurements confirmed roflumilast's mechanism, validating the system.</div></div><div><h3>Significance</h3><div>This strategy eliminates HDCO loss via on-chip integration of all procedures, resolving the conflict between limited samples and multi-condition tests. It improves tracking accuracy by avoiding organoid displacement/overlapping, overcoming multi-well-plate limitations. The system's low cost and high reproducibility accelerate drug screening (e.g., roflumilast) and enable precise study of cilia physiology, laying a key technical foundation for cilia-related disease research and clinical translation.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344967"},"PeriodicalIF":6.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145748490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01DOI: 10.1016/j.aca.2025.344964
Shujun He , Ronghui Liu , Xinxin Bao , Jiayuan Pang , Xi Zhang , Yue Liu , Wenfu Lin , Jiadun Liu , Tianxin Li , Yao Li , Yi Li , Xinrong Guo
Background
The detection of heavy metal ions has traditionally relied on laboratory-based spectroscopy techniques, which are often time-consuming, complex, and unsuitable for real-time monitoring. To address this challenge, we introduce an aptamer conformation sensing strategy and evaluate its ability to detect cadmium ions. The sensing strategy qualitatively infers the isolation of cadmium ions (Cd2+) by monitoring the docking changes of a single DNA aptamer before and after binding with Cd2+ in the nanopore, and quantitatively determines the concentration by extracting the event count of characteristic events.
Result
Molecular dynamics simulations reveal that Cd2+ binding induces structural reconfiguration, reducing aptamer penetration depth from 5.20 nm (free DNA) to 1.66 nm (DNA_Cd2+). Electrical signatures indicate that unbound aptamers produce sustained current blockades (ΔI/I0 = 42 %), while Cd2+ complexes generate transient signals (ΔI/I0 = 25 %). Dual-parameter analysis (ΔI/I0 vs. Irms) provides precise detection. The sensor achieves a detection limit of 33.8 nM within 10 min and demonstrates >93-fold selectivity over Mn2+, Ca2+, Cu2+, Co2+, Ni2+, and Fe3+. Field validation shows detection accuracy consistent with ICP-MS results. Significance: These findings highlight the potential of nanopore-based aptamer conformation sensing as a powerful platform for real-time, quantitative detection of heavy metal ions in complex environmental matrices.
{"title":"Aptamer conformation sensing in nanopore for on-site quantitative detection of cadmium ions at nanomolar concentrations","authors":"Shujun He , Ronghui Liu , Xinxin Bao , Jiayuan Pang , Xi Zhang , Yue Liu , Wenfu Lin , Jiadun Liu , Tianxin Li , Yao Li , Yi Li , Xinrong Guo","doi":"10.1016/j.aca.2025.344964","DOIUrl":"10.1016/j.aca.2025.344964","url":null,"abstract":"<div><h3>Background</h3><div>The detection of heavy metal ions has traditionally relied on laboratory-based spectroscopy techniques, which are often time-consuming, complex, and unsuitable for real-time monitoring. To address this challenge, we introduce an aptamer conformation sensing strategy and evaluate its ability to detect cadmium ions. The sensing strategy qualitatively infers the isolation of cadmium ions (Cd<sup>2+</sup>) by monitoring the docking changes of a single DNA aptamer before and after binding with Cd<sup>2+</sup> in the nanopore, and quantitatively determines the concentration by extracting the event count of characteristic events.</div></div><div><h3>Result</h3><div>Molecular dynamics simulations reveal that Cd<sup>2+</sup> binding induces structural reconfiguration, reducing aptamer penetration depth from 5.20 nm (free DNA) to 1.66 nm (DNA_Cd<sup>2+</sup>). Electrical signatures indicate that unbound aptamers produce sustained current blockades (Δ<em>I/I</em><sub><em>0</em></sub> = 42 %), while Cd<sup>2+</sup> complexes generate transient signals (Δ<em>I/I</em><sub><em>0</em></sub> = 25 %). Dual-parameter analysis (Δ<em>I/I</em><sub><em>0</em></sub> vs. <em>I</em><sub><em>rms</em></sub>) provides precise detection. The sensor achieves a detection limit of 33.8 nM within 10 min and demonstrates >93-fold selectivity over Mn<sup>2+</sup>, Ca<sup>2+</sup>, Cu<sup>2+</sup>, Co<sup>2+</sup>, Ni<sup>2+</sup>, and Fe<sup>3+</sup>. Field validation shows detection accuracy consistent with ICP-MS results. <em>Significance</em>: These findings highlight the potential of nanopore-based aptamer conformation sensing as a powerful platform for real-time, quantitative detection of heavy metal ions in complex environmental matrices.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1384 ","pages":"Article 344964"},"PeriodicalIF":6.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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