Pub Date : 2025-12-04DOI: 10.1016/j.bios.2025.118282
Junyuan Yang, Hao Jiang, Anyi Li, Yulin Deng, Xuefei Lv
The accurate detection of microRNA (miRNA) biomarkers is of paramount importance for early cancer diagnosis. However, it remains challenging due to their low abundance and complex dysregulation patterns in biofluids. Current detection methods often struggle to simultaneously achieve both precision and reliability. To address these limitations, we developed a novel biosensing platform that integrates a primer exchange reaction (PER)-controlled AND logic gate with signal-amplifying “feather-like” nanostructures (FLNs) constructed vis hybridization chain reaction (HCR) for the simultaneous detection of miR-21 and miR-155. This platform employs a core-shell CuS@MIL-101 nanocomposite as a triple-functional signal tag, enabling colorimetric (CL), fluorescent (FL), and photothermal (PT) readouts. The HCR-constructed FLNs provides a high density of binding sites for the signal tags, facilitating cascade signal amplification with up to 2.7-fold, 4.7-fold and 2.7-fold enhancement in three modes, respectively. The PER-mediated logic operation ensures that the signal probe assembly is triggered only in the simultaneous presence of both target miRNAs, enabling attomole-level detection with excellent specificity. The biosensor showed high robustness in spiked human serum, underscoring its strong potential for clinical miRNA analysis and cancer diagnostics.
{"title":"Self-assembled “feather-like” CuS@MIL-101 nanostructure for CL-FL-PT triple-modal signal amplification: PER-controlled AND logic gate detection of miR-21&miR-155","authors":"Junyuan Yang, Hao Jiang, Anyi Li, Yulin Deng, Xuefei Lv","doi":"10.1016/j.bios.2025.118282","DOIUrl":"10.1016/j.bios.2025.118282","url":null,"abstract":"<div><div>The accurate detection of microRNA (miRNA) biomarkers is of paramount importance for early cancer diagnosis. However, it remains challenging due to their low abundance and complex dysregulation patterns in biofluids. Current detection methods often struggle to simultaneously achieve both precision and reliability. To address these limitations, we developed a novel biosensing platform that integrates a primer exchange reaction (PER)-controlled AND logic gate with signal-amplifying “feather-like” nanostructures (FLNs) constructed vis hybridization chain reaction (HCR) for the simultaneous detection of miR-21 and miR-155. This platform employs a core-shell CuS@MIL-101 nanocomposite as a triple-functional signal tag, enabling colorimetric (CL), fluorescent (FL), and photothermal (PT) readouts. The HCR-constructed FLNs provides a high density of binding sites for the signal tags, facilitating cascade signal amplification with up to 2.7-fold, 4.7-fold and 2.7-fold enhancement in three modes, respectively. The PER-mediated logic operation ensures that the signal probe assembly is triggered only in the simultaneous presence of both target miRNAs, enabling attomole-level detection with excellent specificity. The biosensor showed high robustness in spiked human serum, underscoring its strong potential for clinical miRNA analysis and cancer diagnostics.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118282"},"PeriodicalIF":10.5,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118292
Hyun Jung Hwang , Sumita Subedi , Kishor Khadka , Jae-Seon Lee , Keun-Hyeung Lee
Heparan sulfate (HS) proteoglycans on the cell surface play pivotal roles in cell signaling, angiogenesis, endocytosis, infectious disease progression, and tumor invasion. Real-time detection and quantification of HS in live cells remain challenging due to its structural heterogeneity and probe internalization issues. Here, we report rationally designed fluorescent probes (1–3) incorporating a branched peptide receptor with fatty-acid and/or PEG conjugation for selective targeting of HS proteoglycans. Among them, the PEGylated probe 3 showed stable and selective ratiometric fluorescence responses to HS in RPMI medium, with exclusive cell-surface localization for 120 min. In contrast, the fatty-acid-conjugated probe 2 exhibited partial accumulation at the plasma membrane for approximately 20 min, but was subsequently internalized into the cytoplasm, limiting its cell-surface retention. Notably, probe 3 enabled quantitative monitoring of HS depletion in live cells following inhibition of HS sulfation by siRNA or chemical treatment, as well as after heparinase digestion, while simultaneously attenuating fibroblast growth factor signaling and suppressing cancer cell migration. Furthermore, PEG conjugation reduced nonspecific serum protein binding, thereby enhancing sensitivity and selectivity for HS detection in human blood. These findings demonstrate that PEG conjugation is an effective strategy to optimize fluorescent HS probes by enhancing cell-surface anchoring, prolonging retention, and improving ratiometric responses in both cellular and plasma environments.
{"title":"Optimized fluorescent probes for heparan sulfate sensing in live cells and human blood","authors":"Hyun Jung Hwang , Sumita Subedi , Kishor Khadka , Jae-Seon Lee , Keun-Hyeung Lee","doi":"10.1016/j.bios.2025.118292","DOIUrl":"10.1016/j.bios.2025.118292","url":null,"abstract":"<div><div>Heparan sulfate (HS) proteoglycans on the cell surface play pivotal roles in cell signaling, angiogenesis, endocytosis, infectious disease progression, and tumor invasion. Real-time detection and quantification of HS in live cells remain challenging due to its structural heterogeneity and probe internalization issues. Here, we report rationally designed fluorescent probes (<strong>1</strong>–<strong>3</strong>) incorporating a branched peptide receptor with fatty-acid and/or PEG conjugation for selective targeting of HS proteoglycans. Among them, the PEGylated probe <strong>3</strong> showed stable and selective ratiometric fluorescence responses to HS in RPMI medium, with exclusive cell-surface localization for 120 min. In contrast, the fatty-acid-conjugated probe <strong>2</strong> exhibited partial accumulation at the plasma membrane for approximately 20 min, but was subsequently internalized into the cytoplasm, limiting its cell-surface retention. Notably, probe <strong>3</strong> enabled quantitative monitoring of HS depletion in live cells following inhibition of HS sulfation by siRNA or chemical treatment, as well as after heparinase digestion, while simultaneously attenuating fibroblast growth factor signaling and suppressing cancer cell migration. Furthermore, PEG conjugation reduced nonspecific serum protein binding, thereby enhancing sensitivity and selectivity for HS detection in human blood. These findings demonstrate that PEG conjugation is an effective strategy to optimize fluorescent HS probes by enhancing cell-surface anchoring, prolonging retention, and improving ratiometric responses in both cellular and plasma environments.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118292"},"PeriodicalIF":10.5,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118294
Xiaoxu Cao , Rongshen Guo , Shen Wang , Xinqi Cai , Rui Shi , Shengkai Li , Jiayu Zeng , Yuqi Cheng , Qian Dong , Long Chen , Changwei Lin , Zhuo Chen
Developing biochemical sensors that emulate the dynamic selectivity, high specificity and matrix tolerance of living systems is a long-standing goal in analytical science. While significant progress has been made with both functionalized nanomaterials and engineered whole-cell sensors, these approaches often remain limited by static, pre-defined recognition and slow, indirect signal readouts, respectively. Here, we introduce a live cell–plasmonic synergistic sensing system integrating the “sieving-enrichment” capability of native, un-engineered living cells with the direct molecular fingerprinting of SERS to overcome these limitations. In this system, plasmonic nanoparticles are internalized to exploit intracellular transport and enrichment pathways, enabling amplified and high-fidelity SERS signals within confined intracellular spaces to obtain direct molecular recognition. The cell membrane functions as a bio-sieve, providing adaptive matrix tolerance by excluding external interferents, while enabling tunable molecular gating through differential analyte permeability or simply modulation of the cell's metabolic state. This deep bio-integration even enables discrimination between structural analogues as well as different binding states of the same biomarker. This versatile system achieves pretreatment-free, matrix-tolerant quantitative analysis from environmental analysis to biomedical diagnostics. This work establishes a generalizable sensing paradigm whose molecular specificity can be readily realized on-demand by selecting cell lines with distinct transport or metabolic machinery.
{"title":"Live cell–plasmonic synergistic “sieving-enrichment” enables signal amplification and tunable molecular gating for versatile sensing in complex matrices","authors":"Xiaoxu Cao , Rongshen Guo , Shen Wang , Xinqi Cai , Rui Shi , Shengkai Li , Jiayu Zeng , Yuqi Cheng , Qian Dong , Long Chen , Changwei Lin , Zhuo Chen","doi":"10.1016/j.bios.2025.118294","DOIUrl":"10.1016/j.bios.2025.118294","url":null,"abstract":"<div><div>Developing biochemical sensors that emulate the dynamic selectivity, high specificity and matrix tolerance of living systems is a long-standing goal in analytical science. While significant progress has been made with both functionalized nanomaterials and engineered whole-cell sensors, these approaches often remain limited by static, pre-defined recognition and slow, indirect signal readouts, respectively. Here, we introduce a live cell–plasmonic synergistic sensing system integrating the “sieving-enrichment” capability of native, un-engineered living cells with the direct molecular fingerprinting of SERS to overcome these limitations. In this system, plasmonic nanoparticles are internalized to exploit intracellular transport and enrichment pathways, enabling amplified and high-fidelity SERS signals within confined intracellular spaces to obtain direct molecular recognition. The cell membrane functions as a bio-sieve, providing adaptive matrix tolerance by excluding external interferents, while enabling tunable molecular gating through differential analyte permeability or simply modulation of the cell's metabolic state. This deep bio-integration even enables discrimination between structural analogues as well as different binding states of the same biomarker. This versatile system achieves pretreatment-free, matrix-tolerant quantitative analysis from environmental analysis to biomedical diagnostics. This work establishes a generalizable sensing paradigm whose molecular specificity can be readily realized on-demand by selecting cell lines with distinct transport or metabolic machinery.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118294"},"PeriodicalIF":10.5,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118291
Zeinab Rahmati, Jahan Bakhsh Raoof
Crimean-Congo hemorrhagic fever (CCHF) is an acute viral zoonosis and is transmitted through the bite of infected ticks, contact with tissue or blood of infected animals, or secretions of infected individuals. Due to its high mortality rate, no availability of effective treatment or vaccine, and high potential for spread, CCHF poses a serious threat to human health. In response to the need to develop rapid and low-cost detection methods, especially in underserved areas, a sensitive electrochemical immunosensor was designed and evaluated for Crimean-Congo hemorrhagic fever virus (CCHFV) detection. This biosensor was fabricated based on carbon nanofibers (CNFs) modified with copper (II) phosphate nanoflowers (Cu3(PO4)2 NFs), which were used as a flexible and conductive platform to immobilize the desired virus antibody for the design of the proposed electrochemical immunosensor. The cathodic peak current of Cu3(PO4)2 NFs in square wave voltammetry (SWV) was used as an analytical signal for the detection and measurement of the virus on the surface of this immunosensor. The results showed that this immunosensor is capable of detecting the virus in the concentration range of 1.00 × 10 to 1.00 × 108 PFU/mL with a limit of detection (LOD) of 3.30 PFU/mL. This system has features such as a wide linear range, high sensitivity, desirable stability and good reproducibility, that its efficiency was also confirmed in spiked human serum samples. This sensor holds promise for the development of portable tools for rapid detection of CCHFV.
{"title":"Electrochemical immunosensor for sensitive detection of Crimean-Congo hemorrhagic fever virus based on a flexible electrode of carbon nanofibers modified with copper (II) phosphate nanoflowers.","authors":"Zeinab Rahmati, Jahan Bakhsh Raoof","doi":"10.1016/j.bios.2025.118291","DOIUrl":"https://doi.org/10.1016/j.bios.2025.118291","url":null,"abstract":"<p><p>Crimean-Congo hemorrhagic fever (CCHF) is an acute viral zoonosis and is transmitted through the bite of infected ticks, contact with tissue or blood of infected animals, or secretions of infected individuals. Due to its high mortality rate, no availability of effective treatment or vaccine, and high potential for spread, CCHF poses a serious threat to human health. In response to the need to develop rapid and low-cost detection methods, especially in underserved areas, a sensitive electrochemical immunosensor was designed and evaluated for Crimean-Congo hemorrhagic fever virus (CCHFV) detection. This biosensor was fabricated based on carbon nanofibers (CNFs) modified with copper (II) phosphate nanoflowers (Cu<sub>3</sub>(PO<sub>4</sub>)<sub>2</sub> NFs), which were used as a flexible and conductive platform to immobilize the desired virus antibody for the design of the proposed electrochemical immunosensor. The cathodic peak current of Cu<sub>3</sub>(PO<sub>4</sub>)<sub>2</sub> NFs in square wave voltammetry (SWV) was used as an analytical signal for the detection and measurement of the virus on the surface of this immunosensor. The results showed that this immunosensor is capable of detecting the virus in the concentration range of 1.00 × 10 to 1.00 × 10<sup>8</sup> PFU/mL with a limit of detection (LOD) of 3.30 PFU/mL. This system has features such as a wide linear range, high sensitivity, desirable stability and good reproducibility, that its efficiency was also confirmed in spiked human serum samples. This sensor holds promise for the development of portable tools for rapid detection of CCHFV.</p>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"118291"},"PeriodicalIF":10.5,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145712695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118284
Yani Liu , Chenrun Zhang , Xueting Wang , Yiwen Shao , Xinyi Cai , Huan Liu , Hai-Liang Zhu , Xinhua Liu , Yong Qian , Xueao Wang
Lysosomal acidification deficits are increasingly recognized as early events in Alzheimer's disease (AD), yet tools for real-time pH monitoring remain limited. Here, we report AHP, a pH-sensitive fluorescent probe with rapid response (ΔpH ≥0.2), high lysosomal specificity, ultra-large Stokes shift (>240 nm), and excellent photostability for long-term imaging. AHP enabled visualization of biphasic Aβ42-induced lysosomal pH dysregulation—initial hyperacidification followed by pathological alkalinization—directly linking acidification failure to impaired Aβ42 clearance. Dual-channel live imaging showed that reduced lysosomal acidity disrupts Aβ42 degradation, leading to cytoplasmic accumulation and increased toxicity.Using AHP-based high-throughput screening, we identified protocatechuic aldehyde as a lysosomal acidification enhancer and validated its efficacy in restoring autophagic flux. As the probe to achieve super-resolution tracking of lysosomal pH in neurodegeneration, AHP connects pH dynamics with AD pathogenesis and offers a powerful tool for mechanistic insight and drug discovery.This work establishes lysosomal pH homeostasis as a critical target in early AD intervention and highlights the broad applicability of AHP in aging-related disorders.
{"title":"Ultra-large stokes shift probe enables SIM super-resolution tracking of lysosomal acidification impairment in Aβ trafficking dysregulation during Alzheimer's disease","authors":"Yani Liu , Chenrun Zhang , Xueting Wang , Yiwen Shao , Xinyi Cai , Huan Liu , Hai-Liang Zhu , Xinhua Liu , Yong Qian , Xueao Wang","doi":"10.1016/j.bios.2025.118284","DOIUrl":"10.1016/j.bios.2025.118284","url":null,"abstract":"<div><div>Lysosomal acidification deficits are increasingly recognized as early events in Alzheimer's disease (AD), yet tools for real-time pH monitoring remain limited. Here, we report <strong>AHP</strong>, a pH-sensitive fluorescent probe with rapid response (ΔpH ≥0.2), high lysosomal specificity, ultra-large Stokes shift (>240 nm), and excellent photostability for long-term imaging. <strong>AHP</strong> enabled visualization of biphasic Aβ42-induced lysosomal pH dysregulation—initial hyperacidification followed by pathological alkalinization—directly linking acidification failure to impaired Aβ42 clearance. Dual-channel live imaging showed that reduced lysosomal acidity disrupts Aβ42 degradation, leading to cytoplasmic accumulation and increased toxicity.Using <strong>AHP</strong>-based high-throughput screening, we identified protocatechuic aldehyde as a lysosomal acidification enhancer and validated its efficacy in restoring autophagic flux. As the probe to achieve super-resolution tracking of lysosomal pH in neurodegeneration, <strong>AHP</strong> connects pH dynamics with AD pathogenesis and offers a powerful tool for mechanistic insight and drug discovery.This work establishes lysosomal pH homeostasis as a critical target in early AD intervention and highlights the broad applicability of <strong>AHP</strong> in aging-related disorders.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118284"},"PeriodicalIF":10.5,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Glutathione (GSH) and H2O2 are important biomarkers in oxidative stress management, and their imbalance is linked to cancer, neurological illnesses, and metabolic diseases. However, interference from structurally identical antioxidants makes it difficult to detect them selectively and sensitively. To overcome this problem, we developed a defect-engineered Pd-ReS2-MXene nanozyme with improved peroxidase (POD)-mimetic activity for rapid and accurate biomolecular sensing. Atomically dispersed Pd nanoparticles on ReS2-MXene nanosheets, produced through controlled chemical reduction, prove higher catalytic efficiency than standard POD-like catalysts. Compared to Pd-ReS2 (674.8 U mg−1) and Pd-MXene (897.2 U mg−1), the nanozyme exhibits POD-like activity of 1377.6 U mg−1. For 3,3′,5,5′-tetramethylbenzidine (TMB), Michaelis-Menten kinetics show increased catalytic activity with a Vmax of 11.22 μM s−1 and a Km of 0.112 mM. Colorimetric experiments show that Pd-ReS2-MXene catalyzes the H2O2-mediated oxidation of TMB, resulting in a blue product. When GSH reverses the process, TMB reverts to its colorless state. The nanozyme has linear detection ranges of 1–500 nM for H2O2 and 0.05–110 μM for GSH, with detection limits of 0.43 and 1.2 nM, respectively. A paper-based analytical device integrated with a smartphone analyzer allows in situ H2O2 quantification in live cells. GSH detection in serum, plasma, and murine liver tissue lysates confirmed recoveries of 98.4–104 % with low relative standard deviations (1.4–3.4 %) and minimal interference from other amino acids. Pd-ReS2-MXene presents a scalable nanozyme platform for biomedical applications such as disease monitoring and therapeutic interventions, with outstanding reusability, stability, and potential for machine learning-assisted real-time sensing.
谷胱甘肽（GSH）和H2O2是氧化应激管理中的重要生物标志物，它们的失衡与癌症、神经系统疾病和代谢疾病有关。然而，结构相同的抗氧化剂的干扰使它们难以选择性和敏感地检测。为了克服这一问题，我们开发了一种缺陷工程的Pd-ReS2-MXene纳米酶，该酶具有改进的过氧化物酶（POD）模拟活性，用于快速准确的生物分子传感。通过控制化学还原法制备的ReS2-MXene纳米片上原子分散的Pd纳米颗粒，证明了比标准类pod催化剂更高的催化效率。与Pd-ReS2 （674.8 U mg-1）和Pd-MXene （897.2 U mg-1）相比，纳米酶表现出1377.6 U mg-1的pod样活性。对于3,3',5,5'-四甲基联苯胺（TMB）， Michaelis-Menten动力学表现出较高的催化活性，Vmax为11.22 μM s-1， Km为0.112 mM。比色实验表明，Pd-ReS2-MXene催化h2o2介导的TMB氧化反应生成蓝色产物。当谷胱甘肽逆转这一过程时，TMB恢复到无色状态。该酶对H2O2的检测范围为1 ~ 500 nM，对GSH的检测范围为0.05 ~ 110 μM，检出限分别为0.43 nM和1.2 nM。与智能手机分析仪集成的基于纸张的分析设备允许在活细胞中进行原位H2O2定量。血清、血浆和小鼠肝组织裂解物中谷胱甘肽的检测证实回收率为98.4- 104%，相对标准偏差低（1.4- 3.4%），其他氨基酸的干扰最小。Pd-ReS2-MXene提供了一个可扩展的纳米酶平台，用于生物医学应用，如疾病监测和治疗干预，具有出色的可重用性，稳定性和机器学习辅助实时传感的潜力。
{"title":"An AI-assisted smartphone platform for H2O2 and glutathione detection using a dual-functional Pd-ReS2-MXene nanoprobes","authors":"Ezhil Vilian , Jungeun Ahn , Ali Mohammadi , Saeed Reza Hormozi Jangi , Leila Mohammadzadeh , Jaebum Choo , Yun Suk Huh , Young-Kyu Han","doi":"10.1016/j.bios.2025.118287","DOIUrl":"10.1016/j.bios.2025.118287","url":null,"abstract":"<div><div>Glutathione (GSH) and H<sub>2</sub>O<sub>2</sub> are important biomarkers in oxidative stress management, and their imbalance is linked to cancer, neurological illnesses, and metabolic diseases. However, interference from structurally identical antioxidants makes it difficult to detect them selectively and sensitively. To overcome this problem, we developed a defect-engineered Pd-ReS<sub>2</sub>-MXene nanozyme with improved peroxidase (POD)-mimetic activity for rapid and accurate biomolecular sensing. Atomically dispersed Pd nanoparticles on ReS<sub>2</sub>-MXene nanosheets, produced through controlled chemical reduction, prove higher catalytic efficiency than standard POD-like catalysts. Compared to Pd-ReS<sub>2</sub> (674.8 U mg<sup>−1</sup>) and Pd-MXene (897.2 U mg<sup>−1</sup>), the nanozyme exhibits POD-like activity of 1377.6 U mg<sup>−1</sup>. For 3,3′,5,5′-tetramethylbenzidine (TMB), Michaelis-Menten kinetics show increased catalytic activity with a V<sub>max</sub> of 11.22 μM s<sup>−1</sup> and a K<sub>m</sub> of 0.112 mM. Colorimetric experiments show that Pd-ReS<sub>2</sub>-MXene catalyzes the H<sub>2</sub>O<sub>2</sub>-mediated oxidation of TMB, resulting in a blue product. When GSH reverses the process, TMB reverts to its colorless state. The nanozyme has linear detection ranges of 1–500 nM for H<sub>2</sub>O<sub>2</sub> and 0.05–110 μM for GSH, with detection limits of 0.43 and 1.2 nM, respectively. A paper-based analytical device integrated with a smartphone analyzer allows in situ H<sub>2</sub>O<sub>2</sub> quantification in live cells. GSH detection in serum, plasma, and murine liver tissue lysates confirmed recoveries of 98.4–104 % with low relative standard deviations (1.4–3.4 %) and minimal interference from other amino acids. Pd-ReS<sub>2</sub>-MXene presents a scalable nanozyme platform for biomedical applications such as disease monitoring and therapeutic interventions, with outstanding reusability, stability, and potential for machine learning-assisted real-time sensing.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118287"},"PeriodicalIF":10.5,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145686623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Electrochemiluminescence (ECL) has evolved from a simple optical detection method into a powerful probe for interfacial reaction kinetics. By converting electrical signals into optical signals with high temporal and spatial resolution, ECL provides new insights into intermediate dynamic processes such as electron transfer and intermediate formation during the reaction process. This review systematically summarizes and expounds the latest progress in the application of ECL in cross-reaction systems, including oxygen reduction, oxygen evolution, hydrogen evolution and metal ion-related processes. Studies have shown that ECL can be used as a kinetic probe for real-time monitoring of pathways and intermediates that cannot be obtained by conventional electrochemistry. In addition, we highlight its increasingly important role in catalyst evaluation, reaction process monitoring, and single-entity analysis. Finally, we discuss the current challenges and propose future directions, aiming to use ECL as a universal method to decipher the reaction mechanisms at the electrochemical interface.
{"title":"From detection to mechanistic insight: Electrochemiluminescence as a universal platform for reaction kinetics.","authors":"Zhizhi Xiang, Chuhang Wei, Shu Zhu, Shan Liu, Renjun Zhang, Xuehao Tong, Zixu Wang, Jiaxin Deng, Xiang Zhang, Lingfeng Zhong, Ruini Sun, Yahan Hou, Cheng Jiang, Yuchan Zhang, Yang Luo, Guangchao Zang","doi":"10.1016/j.bios.2025.118279","DOIUrl":"https://doi.org/10.1016/j.bios.2025.118279","url":null,"abstract":"<p><p>Electrochemiluminescence (ECL) has evolved from a simple optical detection method into a powerful probe for interfacial reaction kinetics. By converting electrical signals into optical signals with high temporal and spatial resolution, ECL provides new insights into intermediate dynamic processes such as electron transfer and intermediate formation during the reaction process. This review systematically summarizes and expounds the latest progress in the application of ECL in cross-reaction systems, including oxygen reduction, oxygen evolution, hydrogen evolution and metal ion-related processes. Studies have shown that ECL can be used as a kinetic probe for real-time monitoring of pathways and intermediates that cannot be obtained by conventional electrochemistry. In addition, we highlight its increasingly important role in catalyst evaluation, reaction process monitoring, and single-entity analysis. Finally, we discuss the current challenges and propose future directions, aiming to use ECL as a universal method to decipher the reaction mechanisms at the electrochemical interface.</p>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"118279"},"PeriodicalIF":10.5,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145712701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118283
Luca Guagneli , Eszter Supala , Tommi Palomäki , Alejandro García Pérez , Edward Hæggström , Johan Bobacka
Enzymatic electrochemical biosensors are a cornerstone technology in enabling further advancements in the field of Continuous Glucose Monitoring (CGM). Pulsed amperometric methods improve the sensitivity and accuracy of electrochemical biosensors. The literature shows that pulsed amperometry increases the sensitivity of enzymatic glucose biosensors based on Prussian Blue (PB). However, the underlying mechanism responsible for this improvement is poorly understood, which impedes further development of this promising measurement method. The present work elucidates the role of the spontaneous reaction between hydrogen peroxide (H2O2) and Prussian White (PW) in the sensitivity improvement observed with pulsed amperometry. A charged working electrode (WE) containing PW can catalyze the H2O2 reduction in the open-circuit regime (OCP). The consumption of H2O2 over a 30-min contact at OCP was 65 % at a PW WE, compared to 13 % at a PB WE. This spontaneous process is associated with a partial discharge of the WE (PW → PB) between the amperometric pulses. The subsequent re-charging (PB → PW) yields the current amplification observed with pulsed amperometry. Based on this consideration, we developed and validated a model for glucose quantification using pulsed amperometry that considers the spontaneous reaction of H2O2 with PW. The model achieves 0.998 determination coefficient between glucose concentration and four analytical signals. The insights presented in this work support the optimization and development of the pulsed amperometric detection method in enzymatic glucose biosensors. Additionally, this work advances the understanding of H2O2 detection at PB-based sensors, and contributes to the development of precise and accurate enzymatic glucose biosensors.
{"title":"Use of Prussian Blue pseudocapacitive properties to amplify the pulsed amperometric readout of biosensors","authors":"Luca Guagneli , Eszter Supala , Tommi Palomäki , Alejandro García Pérez , Edward Hæggström , Johan Bobacka","doi":"10.1016/j.bios.2025.118283","DOIUrl":"10.1016/j.bios.2025.118283","url":null,"abstract":"<div><div>Enzymatic electrochemical biosensors are a cornerstone technology in enabling further advancements in the field of Continuous Glucose Monitoring (CGM). Pulsed amperometric methods improve the sensitivity and accuracy of electrochemical biosensors. The literature shows that pulsed amperometry increases the sensitivity of enzymatic glucose biosensors based on Prussian Blue (PB). However, the underlying mechanism responsible for this improvement is poorly understood, which impedes further development of this promising measurement method. The present work elucidates the role of the spontaneous reaction between hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) and Prussian White (PW) in the sensitivity improvement observed with pulsed amperometry. A charged working electrode (WE) containing PW can catalyze the H<sub>2</sub>O<sub>2</sub> reduction in the open-circuit regime (OCP). The consumption of H<sub>2</sub>O<sub>2</sub> over a 30-min contact at OCP was 65 % at a PW WE, compared to 13 % at a PB WE. This spontaneous process is associated with a partial discharge of the WE (PW → PB) between the amperometric pulses. The subsequent re-charging (PB → PW) yields the current amplification observed with pulsed amperometry. Based on this consideration, we developed and validated a model for glucose quantification using pulsed amperometry that considers the spontaneous reaction of H<sub>2</sub>O<sub>2</sub> with PW. The model achieves 0.998 determination coefficient between glucose concentration and four analytical signals. The insights presented in this work support the optimization and development of the pulsed amperometric detection method in enzymatic glucose biosensors. Additionally, this work advances the understanding of H<sub>2</sub>O<sub>2</sub> detection at PB-based sensors, and contributes to the development of precise and accurate enzymatic glucose biosensors.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118283"},"PeriodicalIF":10.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691607","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118286
Chengying Zhang , Chao Guan , Qian Liu , Hongbao Fang , Wenjing Song , Xiuzhi Yang , Shankun Yao , Peixue Ling , Xintian Shao , Zhi Su , Yuncong Chen
Unraveling lipid droplets (LDs)-mitochondria-lysosomes crosstalk is vital for decoding cellular states, yet commercial probes suffer from spectral bleed-through, limiting super-resolution imaging. To address this, we innovatively synthesized BDPQ, a LD-targeting fluorescent probe with narrow emission, low toxicity, high cell permeability, and superior photostability—overcoming the flaws of commercial BODIPY dyes. Combining structured illumination microscopy (SIM) with a custom algorithm-assisted analysis, we developed a unique 3D tri-color super-resolution technique, enabling precise quantification of lipophagy, mitophagy, and coordinated lipophagy and mitophagy (named coLipo-Mitophagy) in single cells. Results revealed starvation-induced low-level coLipo-Mitophagy sustains cell viability, while mitochondrial damage drives high-level autophagy and apoptosis. Notably, 2000 kDa hyaluronic acid (HA) was identified as a potential inducer, significantly enhancing coLipo-Mitophagy and triggering LD depletion/apoptosis. This study provides a novel bleed-through-free tool for autophagy-related drug evaluation and inducer discovery.
{"title":"Super-resolution imaging of multiple organelle interactions between lysosome-lipid droplets-mitochondria","authors":"Chengying Zhang , Chao Guan , Qian Liu , Hongbao Fang , Wenjing Song , Xiuzhi Yang , Shankun Yao , Peixue Ling , Xintian Shao , Zhi Su , Yuncong Chen","doi":"10.1016/j.bios.2025.118286","DOIUrl":"10.1016/j.bios.2025.118286","url":null,"abstract":"<div><div>Unraveling lipid droplets (LDs)-mitochondria-lysosomes crosstalk is vital for decoding cellular states, yet commercial probes suffer from spectral bleed-through, limiting super-resolution imaging. To address this, we innovatively synthesized BDPQ, a LD-targeting fluorescent probe with narrow emission, low toxicity, high cell permeability, and superior photostability—overcoming the flaws of commercial BODIPY dyes. Combining structured illumination microscopy (SIM) with a custom algorithm-assisted analysis, we developed a unique 3D tri-color super-resolution technique, enabling precise quantification of lipophagy, mitophagy, and coordinated lipophagy and mitophagy (named coLipo-Mitophagy) in single cells. Results revealed starvation-induced low-level coLipo-Mitophagy sustains cell viability, while mitochondrial damage drives high-level autophagy and apoptosis. Notably, 2000 kDa hyaluronic acid (HA) was identified as a potential inducer, significantly enhancing coLipo-Mitophagy and triggering LD depletion/apoptosis. This study provides a novel bleed-through-free tool for autophagy-related drug evaluation and inducer discovery.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118286"},"PeriodicalIF":10.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145691548","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","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.bios.2025.118280
Weixue Xu , Yan Lin , Zena Huang , Yanyan Li , Yao Lu , Meiru Liu , Sicheng Cui , Tenghua Zhang , Nan Shi , Yan Sheng , Jiaming Hu
Early diagnosis of breast cancer is critical for improving prognosis, but traditional methods have limitations. Herein, we propose an SPC-CRISPR system for the sensitive and specific detection of multiple breast cancer exosomal proteins without prior exosome isolation. This system couples CRISPR system with an enzyme-free amplification method to achieve dual-signal amplification. SPC-CRISPR is based on a split proximity circuit (SPC) that triggers catalytic hairpin assembly (CHA), converting protein signals on the surface of exosomes into nucleic acid signals, and the CRISPR-Cas12a system enabling further signal amplification and output. The system targets phosphatidylserine (PS), MUC1, and EpCAM on exosomes: Tim4-modified magnetic beads capture PS-expressing exosomes, and dual-aptamers recognize MUC1 and EpCAM, enabling SPC assembly and subsequent amplification. In buffer and cell-derived exosomes, the SPC-CRISPR system showed a detection limit of 10 particles/μL (R2 = 0.990). Clinical tests utilizing merely 1 μL of serum samples successfully distinguished breast cancer patients from healthy donors (AUC = 0.9778, accuracy = 91.23 %), detected stage 0 breast cancer patients against healthy controls (accuracy = 92.59 %), and differentiated metastatic from non-metastatic cases (p < 0.001). The combination of high sensitivity, minimal sample requirements, and an exosome isolation-free workflow positions the SPC-CRISPR system as a promising tool for the clinical early detection and classification of breast cancer, with broader applicability to other cancers by swapping the corresponding aptamers.
{"title":"Split proximity circuit initiated CRISPR-Cas12a system profiling exosomal surface proteins for early cancer detection","authors":"Weixue Xu , Yan Lin , Zena Huang , Yanyan Li , Yao Lu , Meiru Liu , Sicheng Cui , Tenghua Zhang , Nan Shi , Yan Sheng , Jiaming Hu","doi":"10.1016/j.bios.2025.118280","DOIUrl":"10.1016/j.bios.2025.118280","url":null,"abstract":"<div><div>Early diagnosis of breast cancer is critical for improving prognosis, but traditional methods have limitations. Herein, we propose an SPC-CRISPR system for the sensitive and specific detection of multiple breast cancer exosomal proteins without prior exosome isolation. This system couples CRISPR system with an enzyme-free amplification method to achieve dual-signal amplification. SPC-CRISPR is based on a split proximity circuit (SPC) that triggers catalytic hairpin assembly (CHA), converting protein signals on the surface of exosomes into nucleic acid signals, and the CRISPR-Cas12a system enabling further signal amplification and output. The system targets phosphatidylserine (PS), MUC1, and EpCAM on exosomes: Tim4-modified magnetic beads capture PS-expressing exosomes, and dual-aptamers recognize MUC1 and EpCAM, enabling SPC assembly and subsequent amplification. In buffer and cell-derived exosomes, the SPC-CRISPR system showed a detection limit of 10 particles/μL (R<sup>2</sup> = 0.990). Clinical tests utilizing merely 1 μL of serum samples successfully distinguished breast cancer patients from healthy donors (AUC = 0.9778, accuracy = 91.23 %), detected stage 0 breast cancer patients against healthy controls (accuracy = 92.59 %), and differentiated metastatic from non-metastatic cases (<em>p</em> < 0.001). The combination of high sensitivity, minimal sample requirements, and an exosome isolation-free workflow positions the SPC-CRISPR system as a promising tool for the clinical early detection and classification of breast cancer, with broader applicability to other cancers by swapping the corresponding aptamers.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"295 ","pages":"Article 118280"},"PeriodicalIF":10.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145659920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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