Pub Date : 2025-01-27DOI: 10.1016/j.bios.2025.117202
Wenyan Li, Zihui Liang, Peilin Wang, Zhenrun Li, Qiang Ma
As an emerging class of extended crystalline organic materials, covalent organic framework (COF)-based aggregation-induced emission luminogen (AIE-gen) exhibited encouraging emissive properties. In this work, 4’,4’’,4‴,4‴’-(1,1,2,2-Ethenetetrayl)tetra(4-biphenylcarbaldehyde) (ETBC) as AIEgen was used to prepare AIE-COF (ET-COF-COOH) luminescent nanoprobe. ETBC and 1,3,5-Tris(4-aminophenyl)benzene (TAPB) had an extended π electronic system that allowed electron delocalization and overlapping transport. Because AIEgen-ETBC served as the luminescence center of ET-COF-COOH, the ET-COF-COOH possessed an ideal anodic electrochemiluminescence (ECL) performance. Moreover, due to the surface plasmonic coupling (SPC) effect of the CuS@Ag square-cavity array, the ECL signal of ET-COF-COOH was amplified as 2.8 times. The AIE-COF/CuS@Ag SCH array-based SPC-ECL sensor was used to detect miRNA-124-3p with a wide range of 1 fM-10 nM and LOD of 0.49 fM. Furthermore, the proposed biosensor can effectively distinguish between tumor tissue and adjacent tissue and offer significant potential for advancing glioma diagnosis.
{"title":"CuS@Ag Heterostructure-based Surface Plasmonic Coupling Electrochemiluminescence Sensor for Glioma miRNA-124-3p Detection","authors":"Wenyan Li, Zihui Liang, Peilin Wang, Zhenrun Li, Qiang Ma","doi":"10.1016/j.bios.2025.117202","DOIUrl":"10.1016/j.bios.2025.117202","url":null,"abstract":"<div><div>As an emerging class of extended crystalline organic materials, covalent organic framework (COF)-based aggregation-induced emission luminogen (AIE-gen) exhibited encouraging emissive properties. In this work, 4’,4’’,4‴,4‴’-(1,1,2,2-Ethenetetrayl)tetra(4-biphenylcarbaldehyde) (ETBC) as AIEgen was used to prepare AIE-COF (ET-COF-COOH) luminescent nanoprobe. ETBC and 1,3,5-Tris(4-aminophenyl)benzene (TAPB) had an extended π electronic system that allowed electron delocalization and overlapping transport. Because AIEgen-ETBC served as the luminescence center of ET-COF-COOH, the ET-COF-COOH possessed an ideal anodic electrochemiluminescence (ECL) performance. Moreover, due to the surface plasmonic coupling (SPC) effect of the CuS@Ag square-cavity array, the ECL signal of ET-COF-COOH was amplified as 2.8 times. The AIE-COF/CuS@Ag SCH array-based SPC-ECL sensor was used to detect miRNA-124-3p with a wide range of 1 fM-10 nM and LOD of 0.49 fM. Furthermore, the proposed biosensor can effectively distinguish between tumor tissue and adjacent tissue and offer significant potential for advancing glioma diagnosis.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"274 ","pages":"Article 117202"},"PeriodicalIF":10.7,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143062586","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 : 2025-01-25DOI: 10.1016/j.bios.2025.117180
Daniel Quesada-González , Arben Merkoçi
Quantum dots (QDs) are the smallest nanomaterials (2–10 nm), with unique optical and electronic properties. Thanks to these properties, QDs have been standing during the last years as signal tags for different applications, including bioimaging, fluorescent biosensors and electrochemical assays. In this review, we explore the current state-of-the art on these nanomaterials, differentiating them between semiconductor and carbon-based QDs. Also, the review focuses on their unique advantages as transducers in different biosensing platforms.
{"title":"Quantum dots for biosensing: Classification and applications","authors":"Daniel Quesada-González , Arben Merkoçi","doi":"10.1016/j.bios.2025.117180","DOIUrl":"10.1016/j.bios.2025.117180","url":null,"abstract":"<div><div>Quantum dots (QDs) are the smallest nanomaterials (2–10 nm), with unique optical and electronic properties. Thanks to these properties, QDs have been standing during the last years as signal tags for different applications, including bioimaging, fluorescent biosensors and electrochemical assays. In this review, we explore the current state-of-the art on these nanomaterials, differentiating them between semiconductor and carbon-based QDs. Also, the review focuses on their unique advantages as transducers in different biosensing platforms.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117180"},"PeriodicalIF":10.7,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143045315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-25DOI: 10.1016/j.bios.2025.117197
Haote Han , Xia Ying , Qiaoqiao Chen , Jiaru Fang , Dongxin Xu , Xuelian Lyu , Jilin Zheng , Ling Zou , Qiong Luo , Ning Hu
Preterm birth (PTB) remains a leading cause of neonatal morbidity and mortality, with inflammation-induced PTB posing a significant challenge due to its complex pathophysiology. To address this, we developed an in vitro platform utilizing hTERT-immortalized human myometrial (hTERT-HM) cells integrated with a multielectrode array (MEA) biosensing system and optical calcium imaging. Compared to primary uterine myometrial cells, hTERT-HM cells exhibit superior reproducibility, high scalability, and convenient manipulation, facilitating the consistent and large-scale investigations. This advanced system facilitates simultaneous real-time monitoring of electrophysiological activity and intracellular calcium transient, providing detailed insights into uterine cell behavior during inflammatory PTB. Our study revealed that oxytocin (OT) induces regular contractions in hTERT-HM cells, and the synergistic effect of OT and lipopolysaccharide (LPS) disrupts electrophysiological patterns and calcium signaling, closely mimicking the pathophysiology of inflammation-induced PTB. Meanwhile, magnesium sulfate is validated to effectively suppress OT-induced calcium release and mitigate LPS-triggered irregular electrophysiological signals. By integrating advanced biosensing technologies and advantages of hTERT-HM cells, this platform offers a reliable, reproducible model to investigate the mechanisms of inflammation-driven PTB and further develop targeted therapeutic interventions.
{"title":"Monitoring of inflammatory preterm responses via myometrial cell based multimodal electrophysiological and optical biosensing platform","authors":"Haote Han , Xia Ying , Qiaoqiao Chen , Jiaru Fang , Dongxin Xu , Xuelian Lyu , Jilin Zheng , Ling Zou , Qiong Luo , Ning Hu","doi":"10.1016/j.bios.2025.117197","DOIUrl":"10.1016/j.bios.2025.117197","url":null,"abstract":"<div><div>Preterm birth (PTB) remains a leading cause of neonatal morbidity and mortality, with inflammation-induced PTB posing a significant challenge due to its complex pathophysiology. To address this, we developed an <em>in vitro</em> platform utilizing hTERT-immortalized human myometrial (hTERT-HM) cells integrated with a multielectrode array (MEA) biosensing system and optical calcium imaging. Compared to primary uterine myometrial cells, hTERT-HM cells exhibit superior reproducibility, high scalability, and convenient manipulation, facilitating the consistent and large-scale investigations. This advanced system facilitates simultaneous real-time monitoring of electrophysiological activity and intracellular calcium transient, providing detailed insights into uterine cell behavior during inflammatory PTB. Our study revealed that oxytocin (OT) induces regular contractions in hTERT-HM cells, and the synergistic effect of OT and lipopolysaccharide (LPS) disrupts electrophysiological patterns and calcium signaling, closely mimicking the pathophysiology of inflammation-induced PTB. Meanwhile, magnesium sulfate is validated to effectively suppress OT-induced calcium release and mitigate LPS-triggered irregular electrophysiological signals. By integrating advanced biosensing technologies and advantages of hTERT-HM cells, this platform offers a reliable, reproducible model to investigate the mechanisms of inflammation-driven PTB and further develop targeted therapeutic interventions.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"274 ","pages":"Article 117197"},"PeriodicalIF":10.7,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143057635","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 : 2025-01-25DOI: 10.1016/j.bios.2025.117184
Jiangtao Ning , Yikai Shen , Hongfan Gao, Li Sun, Xuefei Bai, Shijie Jin, Yue Wu, Yanping Sun, Yingchun Xu, Xin Li, Liqiang Pan
Antibody-drug conjugates (ADC) have emerged as an important class of therapeutic agents that combine the target specificity of a monoclonal antibody with the potency of a cytotoxic payload. Despite clinical success, our understanding of receptor endocytosis and ADC toxicity remains limited. Less than 1% of ADCs reach tumors, raising concerns about off-target cytotoxicity. To shed light on these issues, our study introduces a smart antibody-fluorophore conjugate (sAFC) with Cathepsin B dependent Activatable Trigger Fluorophore (CAT-Fluor) to mimic ADC behavior in situ. Using a Cathepsin B-cleavable linker, we linked a Si-rhodamine (SiR) derivative with superior near-infrared emission to antibodies, creating sAFC. Carbamoylation of the primary amino group on SiR is employed to conjugate with the linker and inhibit the electron-push-pull effect of the xanthene skeleton, thus inducing fluorescence quenching. In vitro, the anti-EGFR sAFC emulates ADC metabolism and suggests that specific proteins implicated in endocytosis, like caveolin, significantly influence ADC internalization efficacy, potentially correlating with drug resistance. In vivo studies using sAFC demonstrate that 'passenger ADCs' found in normal tissues release minimal payload, likely elucidating how ADCs mitigate dose-limiting toxicities. Therefore, our sAFC-based strategy, combining CAT-Fluor and targeted interventions, quantitatively and objectively evaluated the impact of various stages and key proteins in the physiological process, spanning from antigen recognition, endocytosis mechanism, to transport and protein hydrolysis, on ADC efficiency. This comprehensive approach lays a mechanistic foundation for advancing ADC research and development, and offers novel insights into tackling ADC efficacy, resistance and potential toxicities from the standpoint of endocytosis mechanisms.
{"title":"Cathepsin B dependent activatable trigger fluorophore (CAT-Fluor) for in situ functional imaging of antibody-drug conjugates","authors":"Jiangtao Ning , Yikai Shen , Hongfan Gao, Li Sun, Xuefei Bai, Shijie Jin, Yue Wu, Yanping Sun, Yingchun Xu, Xin Li, Liqiang Pan","doi":"10.1016/j.bios.2025.117184","DOIUrl":"10.1016/j.bios.2025.117184","url":null,"abstract":"<div><div>Antibody-drug conjugates (ADC) have emerged as an important class of therapeutic agents that combine the target specificity of a monoclonal antibody with the potency of a cytotoxic payload. Despite clinical success, our understanding of receptor endocytosis and ADC toxicity remains limited. Less than 1% of ADCs reach tumors, raising concerns about off-target cytotoxicity. To shed light on these issues, our study introduces a smart antibody-fluorophore conjugate (sAFC) with Cathepsin B dependent Activatable Trigger Fluorophore (CAT-Fluor) to mimic ADC behavior <em>in situ</em>. Using a Cathepsin B-cleavable linker, we linked a Si-rhodamine (SiR) derivative with superior near-infrared emission to antibodies, creating sAFC. Carbamoylation of the primary amino group on SiR is employed to conjugate with the linker and inhibit the electron-push-pull effect of the xanthene skeleton, thus inducing fluorescence quenching. In vitro, the anti-EGFR sAFC emulates ADC metabolism and suggests that specific proteins implicated in endocytosis, like caveolin, significantly influence ADC internalization efficacy, potentially correlating with drug resistance. In vivo studies using sAFC demonstrate that 'passenger ADCs' found in normal tissues release minimal payload, likely elucidating how ADCs mitigate dose-limiting toxicities. Therefore, our sAFC-based strategy, combining CAT-Fluor and targeted interventions, quantitatively and objectively evaluated the impact of various stages and key proteins in the physiological process, spanning from antigen recognition, endocytosis mechanism, to transport and protein hydrolysis, on ADC efficiency. This comprehensive approach lays a mechanistic foundation for advancing ADC research and development, and offers novel insights into tackling ADC efficacy, resistance and potential toxicities from the standpoint of endocytosis mechanisms.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"274 ","pages":"Article 117184"},"PeriodicalIF":10.7,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143062574","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 : 2025-01-23DOI: 10.1016/j.bios.2025.117187
Yuning Zhang , Yanbing Lv , Xinxin Chen , Yingli Ye , Shenping Yu , Ning Li , Hongtao Yao , Zhizeng Wang , Qitong Feng , Ruili Wu , Lin Song Li
Influenza A virus (IAV) and influenza B virus (IBV) with similar symptoms of infection caused a serious disease burden and economic losses in annual epidemic season, so it is important to quickly and accurately detect and distinguish between IAV and IBV during influenza season. Herein, the quantum dot microspheres (QDMS) were synthesized and applied to lateral flow immunoassays (LFIA), and a point-of-care (POC) biosensor that can discriminately and simultaneously diagnose IAV and IBV within 10 min was established. A double-sandwich QDMS nanotags was synthesized by immobilizing hydrophobic quantum dots (QDs) with chemical bonding method on a silica sphere template with an outer silica shell protection showed excellent stability and high fluorescence. The highly sensitive quantitative detection is achieved with the tailored hand-hold detector. The limit of detection (LOD) of the LFIA based QDMS for the simultaneous detection of IAV and IBV were 0.080 ng/mL and 0.096 ng/mL, respectively. The QDMS-LFIA system has excellent specificity and stability for quantitative detection, especially showing good accuracy for clinical samples. Furthermore, the visualization sensitivity of the low-cost biosensor was improved by nearly an order of magnitude than gold nanoparticles (AuNP)-LFIA within shorter detection time. Given the excellent performance, our proposed QDMS-LFIA biosensor can potentially be applied to the POC sensitive discriminate and simultaneous detection of IAV and IBV in epidemic season.
{"title":"Ultrasensitive point-of-care multiplex diagnosis for influenza virus based robust quantum dot microsphere-lateral flow immunoassay","authors":"Yuning Zhang , Yanbing Lv , Xinxin Chen , Yingli Ye , Shenping Yu , Ning Li , Hongtao Yao , Zhizeng Wang , Qitong Feng , Ruili Wu , Lin Song Li","doi":"10.1016/j.bios.2025.117187","DOIUrl":"10.1016/j.bios.2025.117187","url":null,"abstract":"<div><div>Influenza A virus (IAV) and influenza B virus (IBV) with similar symptoms of infection caused a serious disease burden and economic losses in annual epidemic season, so it is important to quickly and accurately detect and distinguish between IAV and IBV during influenza season. Herein, the quantum dot microspheres (QDMS) were synthesized and applied to lateral flow immunoassays (LFIA), and a point-of-care (POC) biosensor that can discriminately and simultaneously diagnose IAV and IBV within 10 min was established. A double-sandwich QDMS nanotags was synthesized by immobilizing hydrophobic quantum dots (QDs) with chemical bonding method on a silica sphere template with an outer silica shell protection showed excellent stability and high fluorescence. The highly sensitive quantitative detection is achieved with the tailored hand-hold detector. The limit of detection (LOD) of the LFIA based QDMS for the simultaneous detection of IAV and IBV were 0.080 ng/mL and 0.096 ng/mL, respectively. The QDMS-LFIA system has excellent specificity and stability for quantitative detection, especially showing good accuracy for clinical samples. Furthermore, the visualization sensitivity of the low-cost biosensor was improved by nearly an order of magnitude than gold nanoparticles (AuNP)-LFIA within shorter detection time. Given the excellent performance, our proposed QDMS-LFIA biosensor can potentially be applied to the POC sensitive discriminate and simultaneous detection of IAV and IBV in epidemic season.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117187"},"PeriodicalIF":10.7,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143051161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-22DOI: 10.1016/j.bios.2025.117186
Wei Li , Shuaijing Wang , Minzhao Lin , Xueying Chen , Jiayue Li , Wanling Cui , Rui Wang
This study presents a dual-mode and regenerated DNA motor powered by exonuclease III (Exo III) for the simultaneous detection of viral gene fragments. The detection methodology is categorized into two distinct operational modes. The first mode emphasizes the simultaneous detection of two viral gene fragments from a specific virus. The presence of both genes triggers the operation of the DNA motor, generating a singular signal output. This mode operates on an “AND” logical mechanism, which enhances the precision of positive case identification. The second mode facilitates the simultaneous detection of three viral gene fragments from three different viruses within a single assay. The presence of these genes activates their respective motors, yielding distinct signal outputs. This mode supports the multiplex detection of three target genes, thereby aiding in the identification of previously uncharacterized viruses infecting patients and alleviating the logistical and financial burdens associated with multiple testing procedures. The detection limit in the “AND” logical mode is at the aM level, while in the multiplex mode, it reaches the fM level, facilitating the sensitive detection of viral gene fragments. The DNA motor can be regenerated by separating and reconstituting the utilized orbits, enabling its reuse for up to seven cycles in the “AND” logical mode and five cycles in the multiplex mode. Accurate diagnoses were achieved for patients exhibiting upper respiratory symptoms. Therefore, the proposed motor offers a novel and regenerative approach for viral gene fragments detection, demonstrating significant promise for application in the clinical diagnosis of viral infectious diseases.
{"title":"Dual-mode, regenerated DNA motor for simultaneous detection of viral gene fragments and diagnosis of infectious disease","authors":"Wei Li , Shuaijing Wang , Minzhao Lin , Xueying Chen , Jiayue Li , Wanling Cui , Rui Wang","doi":"10.1016/j.bios.2025.117186","DOIUrl":"10.1016/j.bios.2025.117186","url":null,"abstract":"<div><div>This study presents a dual-mode and regenerated DNA motor powered by exonuclease III (Exo III) for the simultaneous detection of viral gene fragments. The detection methodology is categorized into two distinct operational modes. The first mode emphasizes the simultaneous detection of two viral gene fragments from a specific virus. The presence of both genes triggers the operation of the DNA motor, generating a singular signal output. This mode operates on an “AND” logical mechanism, which enhances the precision of positive case identification. The second mode facilitates the simultaneous detection of three viral gene fragments from three different viruses within a single assay. The presence of these genes activates their respective motors, yielding distinct signal outputs. This mode supports the multiplex detection of three target genes, thereby aiding in the identification of previously uncharacterized viruses infecting patients and alleviating the logistical and financial burdens associated with multiple testing procedures. The detection limit in the “AND” logical mode is at the aM level, while in the multiplex mode, it reaches the fM level, facilitating the sensitive detection of viral gene fragments. The DNA motor can be regenerated by separating and reconstituting the utilized orbits, enabling its reuse for up to seven cycles in the “AND” logical mode and five cycles in the multiplex mode. Accurate diagnoses were achieved for patients exhibiting upper respiratory symptoms. Therefore, the proposed motor offers a novel and regenerative approach for viral gene fragments detection, demonstrating significant promise for application in the clinical diagnosis of viral infectious diseases.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117186"},"PeriodicalIF":10.7,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035379","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 : 2025-01-22DOI: 10.1016/j.bios.2025.117158
Norh Asmare , A K M Arifuzzman , Ningquan Wang , Mert Boya , Ruxiu Liu , A. Fatih Sarioglu
Since physiological and pathological events change the mechanical properties of cells, tools that rapidly quantify such changes at the single-cell level can advance the utility of cell mechanics as a label-free biomarker. We demonstrate the capability to probe the population-level elastic modulus and fluidity of MDA-MB-231 cells at a throughput of up to 50 cell/second within a portable microchip. Our sensing scheme adapts a code multiplexing scheme to implement a distributed network of sensors throughout the microchip, thereby compressing all sensing events into a single electrical output. To validate our approach, we prepared cell samples whose stiffnesses were manipulated with chemical agents. We confirmed the expected effect of the chemicals agreed with the stiffness measurements reported by our microchip. Such a low-cost electronic assay that rapidly measures mechanical properties enables previously infeasible studies to advance the science of mechanobiology.
{"title":"High throughput cell stiffness measurement via multiplexed impedance sensors","authors":"Norh Asmare , A K M Arifuzzman , Ningquan Wang , Mert Boya , Ruxiu Liu , A. Fatih Sarioglu","doi":"10.1016/j.bios.2025.117158","DOIUrl":"10.1016/j.bios.2025.117158","url":null,"abstract":"<div><div>Since physiological and pathological events change the mechanical properties of cells, tools that rapidly quantify such changes at the single-cell level can advance the utility of cell mechanics as a label-free biomarker. We demonstrate the capability to probe the population-level elastic modulus and fluidity of MDA-MB-231 cells at a throughput of up to 50 cell/second within a portable microchip. Our sensing scheme adapts a code multiplexing scheme to implement a distributed network of sensors throughout the microchip, thereby compressing all sensing events into a single electrical output. To validate our approach, we prepared cell samples whose stiffnesses were manipulated with chemical agents. We confirmed the expected effect of the chemicals agreed with the stiffness measurements reported by our microchip. Such a low-cost electronic assay that rapidly measures mechanical properties enables previously infeasible studies to advance the science of mechanobiology.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117158"},"PeriodicalIF":10.7,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143027471","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 : 2025-01-22DOI: 10.1016/j.bios.2025.117183
Li Zhang , Tong Feng , Qian Liu , Chen Zuo , Yongchang Wu , Huaixin Zhao , Hongyan Yu , Dan Bai , Xiaole Han , Na Yin , Jiu Pu , Yujun Yang , Junjie Li , Jinhong Guo , Shixiong Deng , Guoming Xie
Isothermal amplification-based nucleic acid detection technologies have become rapid and efficient tools for molecular diagnostics. Sequence-specific monitoring methods are crucial for isothermal amplification, as they help identify the occurrence of extended primer dimers, which can lead to false positive results. Fluorescent aptamers are promising tools for real-time monitoring of isothermal amplification but are inherently limited by thermostability. Here, we report an engineered fluorescent DNA aptamer variant, named thermostable Lettuce (TS-Lettuce), with a 5 °C higher melting temperature and 20 times greater fluorescence at 60 °C, ideal for real-time monitoring of sequence-specific isothermal amplification. Using molecular dynamics simulations for structural analyses, we introduced mutations to wild-type Lettuce to redesign the non-core sequences of the aptamer structure for tightly stabilizing its folding, thereby enhancing thermostability. The TS-Lettuce offers greater versatility and ease of design for coupling with isothermal amplification for all-in-one nucleic acid detection. We demonstrated three applications of TS-Lettuce in isothermal amplification: fluorescent turn-off, fluorescent turn-on, and fluorescent aptamer switch, facilitating the sequence-specific detection of nucleic acids. In addition, the results generated by TS-Lettuce are visible to the naked eye, enhancing the utility of isothermal amplification reactions in resource-constrained areas. The thermostable fluorescent DNA aptamers can be further utilized in more isothermal amplification methods.
{"title":"Engineering thermostable fluorescent DNA aptamer for the isothermal amplification of nucleic acids","authors":"Li Zhang , Tong Feng , Qian Liu , Chen Zuo , Yongchang Wu , Huaixin Zhao , Hongyan Yu , Dan Bai , Xiaole Han , Na Yin , Jiu Pu , Yujun Yang , Junjie Li , Jinhong Guo , Shixiong Deng , Guoming Xie","doi":"10.1016/j.bios.2025.117183","DOIUrl":"10.1016/j.bios.2025.117183","url":null,"abstract":"<div><div>Isothermal amplification-based nucleic acid detection technologies have become rapid and efficient tools for molecular diagnostics. Sequence-specific monitoring methods are crucial for isothermal amplification, as they help identify the occurrence of extended primer dimers, which can lead to false positive results. Fluorescent aptamers are promising tools for real-time monitoring of isothermal amplification but are inherently limited by thermostability. Here, we report an engineered fluorescent DNA aptamer variant, named thermostable Lettuce (TS-Lettuce), with a 5 °C higher melting temperature and 20 times greater fluorescence at 60 °C, ideal for real-time monitoring of sequence-specific isothermal amplification. Using molecular dynamics simulations for structural analyses, we introduced mutations to wild-type Lettuce to redesign the non-core sequences of the aptamer structure for tightly stabilizing its folding, thereby enhancing thermostability. The TS-Lettuce offers greater versatility and ease of design for coupling with isothermal amplification for all-in-one nucleic acid detection. We demonstrated three applications of TS-Lettuce in isothermal amplification: fluorescent turn-off, fluorescent turn-on, and fluorescent aptamer switch, facilitating the sequence-specific detection of nucleic acids. In addition, the results generated by TS-Lettuce are visible to the naked eye, enhancing the utility of isothermal amplification reactions in resource-constrained areas. The thermostable fluorescent DNA aptamers can be further utilized in more isothermal amplification methods.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117183"},"PeriodicalIF":10.7,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035494","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}
We report a stand-alone, automated, fully quantitative, and portable Microfluidics Integrated LED-Photodiode (MILP) sensing technology as a new molecular diagnostic platform for rapid point-of-care nucleic acid testing in real-time. The all-in-one device integrates a paper-based assay for nucleic acid purification using a polymer-based membrane filter, in-situ isothermal amplification, dual-mode optical detection, and fully quantitative signal analysis by capturing the photovoltaic response using electrical polarity and photocurrent measurements. Highly selective photovoltaic cut-offs may readily recognize test-gene-specific variations quantitatively without requiring further auxiliary instrumentation. The on-cartridge limit of detection (LoD) showed 10 copies/μL, which could diagnose SARS-CoV-2 samples with high clinical sensitivity (95%) and specificity (100%) with reference to real-time PCR-based gold-standard benchmark. Our findings emphasize the test's unique advantages for intensive health surveillance, enabling early disease screening, precise severity assessment, and real-time tracking of disease progression in resource-limited settings without the need for extensive and expensive laboratory infrastructure.
{"title":"Portable, quantitative, real-time isothermal nucleic acid amplification test using microfluidic device-coupled UV-LED photodiode detector","authors":"Natish kumar , Monika Kumari , Devtulya Chander , Sandeep Dogra , Asha Chaubey , Suman Chakraborty , Ravi Kumar Arun","doi":"10.1016/j.bios.2025.117194","DOIUrl":"10.1016/j.bios.2025.117194","url":null,"abstract":"<div><div>We report a stand-alone, automated, fully quantitative, and portable Microfluidics Integrated LED-Photodiode (MILP) sensing technology as a new molecular diagnostic platform for rapid point-of-care nucleic acid testing in real-time. The all-in-one device integrates a paper-based assay for nucleic acid purification using a polymer-based membrane filter, in-situ isothermal amplification, dual-mode optical detection, and fully quantitative signal analysis by capturing the photovoltaic response using electrical polarity and photocurrent measurements. Highly selective photovoltaic cut-offs may readily recognize test-gene-specific variations quantitatively without requiring further auxiliary instrumentation. The on-cartridge limit of detection (LoD) showed 10 copies/μL, which could diagnose SARS-CoV-2 samples with high clinical sensitivity (95%) and specificity (100%) with reference to real-time PCR-based gold-standard benchmark. Our findings emphasize the test's unique advantages for intensive health surveillance, enabling early disease screening, precise severity assessment, and real-time tracking of disease progression in resource-limited settings without the need for extensive and expensive laboratory infrastructure.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"274 ","pages":"Article 117194"},"PeriodicalIF":10.7,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143171474","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 : 2025-01-22DOI: 10.1016/j.bios.2025.117193
Bingun Habarakadage , Sabari Rajendran , Zhaoyang Ren , Morgan J. Anderson , Jessica Koehne , Lingaraju Gorla , Shunya Morita , Sara Wu , Duy H. Hua , Jun Li
Proteases are overexpressed at various stages of conditions such as cancers and thus can serve as biomarkers for disease diagnosis. Electrochemical techniques to detect the activity of extracellular proteases have gained attraction due to their multiplexing capability. Here we employ an electrochemical approach based on a 3 × 3 gold (Au) microelectrode array (MEA) functionalized with (2-aminoethyl)ferrocene (AEF) tagged specific peptide substrates to monitor cathepsin B (CB) protease activity. Cleavage of these peptide substrates by proteases leads to an exponential decay in the alternating current voltammetry (ACV) signal. The protease activity is represented by the inverse of the decay time constant (1/τ), which is equal to (kcat/KM)[CB] based on the heterogeneous Michaelis-Menton model. However, the thiol/Au chemisorption linking AEF-peptide to gold electrodes is susceptible to interference by the protease activation reagent dithiothreitol (DTT), causing the peptides to desorb from the Au surface during continuous ACV measurement. This induces a false signal decay, masking the protease activity and reducing the sensor sensitivity. To address this, DTT is removed after activating CB using centrifugal filtration while EDTA is incorporated to maintain the enzyme activity. This allows accurate CB proteolysis kinetics and clarifies the roles of EDTA and DTT in activation. The intrinsic substrate-dependent cleavage by CB to three different peptide substrates has been demonstrated with the MEA chip, showcasing the potential for rapid activity profiling of multiple proteases. The study highlights the importance of understanding the interference of active bioreagents to the thiol/Au interface in broad redox-tagged electrochemical biosensors.
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