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Integrated microphotonic platform for highly sensitive virus detection 集成微光子平台,用于高灵敏度病毒检测
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-04-09 DOI: 10.1016/j.snr.2025.100320
A. la Grasta , M.I. Gómez-Gómez , A. Griol , E. Gómez , M. De Carlo , V.M.N. Passaro , A. Martínez , F. Dell’Olio
{"title":"Integrated microphotonic platform for highly sensitive virus detection","authors":"A. la Grasta ,&nbsp;M.I. Gómez-Gómez ,&nbsp;A. Griol ,&nbsp;E. Gómez ,&nbsp;M. De Carlo ,&nbsp;V.M.N. Passaro ,&nbsp;A. Martínez ,&nbsp;F. Dell’Olio","doi":"10.1016/j.snr.2025.100320","DOIUrl":"10.1016/j.snr.2025.100320","url":null,"abstract":"<div><div>Chip-scale micro-photonic biosensors have matured significantly over the past two decades, with various platforms developed for detecting biomarkers, including proteins in biological fluids. The COVID-19 pandemic has further accelerated research in virus detection, leading to the development of resonant and interferometric biosensors for identifying viruses like SARS-CoV-2. Despite these advancements, there remains a need for technical solutions that ensure point-of-care features such as low cost, rapid response, and low limit-of-detection (LoD). This paper presents an integrated silicon nitride microphotonic platform based on ring resonator technology, specifically designed for highly sensitive virus detection. The sensors, fabricated using strip and slot waveguides, are tested under flow conditions using ethanol and BSA-antiBSA solutions to evaluate bulk and surface sensitivity. The results indicate that the slot waveguide-based ring resonators exhibit the highest sensitivity. The platform uses functionalization protocols to immobilize specific antibodies for detecting SARS-CoV-2. The sensor successfully identifies the target analyte at concentrations as low as 1.5 nM, with a LoD of 36 pM using in situ functionalization, underscoring the importance of antibody orientation through an intermediate protein A layer. These results demonstrate the platform’s potential for rapid and sensitive virus diagnosis and represent a significant step toward the development of portable, point-of-care diagnostic technologies for infectious disease monitoring.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100320"},"PeriodicalIF":6.5,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143820877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Ultra-sensitive aptameric field-effect transistor for cortisol sensing 用于皮质醇传感的超灵敏适体场效应晶体管
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-04-08 DOI: 10.1016/j.snr.2025.100324
Thi Thanh Ha Nguyen , Cong Minh Nguyen , Minh Anh Huynh , Quang Thang Trinh , Philip Tanner , Sven Ingebrandt , Xuan Thang Vu , Tuan-Khoa Nguyen , Nam-Trung Nguyen
{"title":"Ultra-sensitive aptameric field-effect transistor for cortisol sensing","authors":"Thi Thanh Ha Nguyen ,&nbsp;Cong Minh Nguyen ,&nbsp;Minh Anh Huynh ,&nbsp;Quang Thang Trinh ,&nbsp;Philip Tanner ,&nbsp;Sven Ingebrandt ,&nbsp;Xuan Thang Vu ,&nbsp;Tuan-Khoa Nguyen ,&nbsp;Nam-Trung Nguyen","doi":"10.1016/j.snr.2025.100324","DOIUrl":"10.1016/j.snr.2025.100324","url":null,"abstract":"<div><div>Existing diagnostic methods for major depressive disorders, emerging mental disorders globally, are restrictive and inaccurate due to the lack of evidence-based assessments. Recent developments incorporating advanced material, micro/nano manufacturing and biology enable new clinical pathways using biosensors that can provide new opportunities for the diagnosis of mental condition from an early developmental stage. In this work, we present an ultra-sensitive lab-on-chip biosensor platform that is capable of detecting cortisol, a key biomarker for depression in physiological fluid. Utilizing advanced microfabrication techniques, our biosensor achieves a compact footprint of 5.0 mm by 2.5 mm, making it suitable for integration into portable and wearable devices. The device exhibits an ultra-low detection limit of 1 fM and a wide dynamic detection range spanning from 1 fM to 1 µM, covering the concentration present in physiological body fluids of human. Our finding shows a great potential for the development of accurate, and point-of-care monitoring system for early detection of depression disorders to advance the mental health diagnosis.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100324"},"PeriodicalIF":6.5,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Argonaute-based dual lateral flow test for SARS-CoV-2 and influenza a virus 基于 Argonaute 的 SARS-CoV-2 和甲型流感病毒双侧流试验
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-04-02 DOI: 10.1016/j.snr.2025.100321
Xingyu Ye , Zhonglei Li , Zhengwei Fang , Nan Shen , Yani Zhang , Hanming Wang , Yi Liu , Yuanjie Zhou , Shanshan Fan , Peng Sun , Yue Zhong , Qian Liu , Yan Feng
{"title":"Argonaute-based dual lateral flow test for SARS-CoV-2 and influenza a virus","authors":"Xingyu Ye ,&nbsp;Zhonglei Li ,&nbsp;Zhengwei Fang ,&nbsp;Nan Shen ,&nbsp;Yani Zhang ,&nbsp;Hanming Wang ,&nbsp;Yi Liu ,&nbsp;Yuanjie Zhou ,&nbsp;Shanshan Fan ,&nbsp;Peng Sun ,&nbsp;Yue Zhong ,&nbsp;Qian Liu ,&nbsp;Yan Feng","doi":"10.1016/j.snr.2025.100321","DOIUrl":"10.1016/j.snr.2025.100321","url":null,"abstract":"<div><div>The lateral flow test (LFT) is crucial for community screening and early warning of infectious pathogens due to its portability and simplicity. However, antigen-based LFTs have relatively low sensitivity, while nucleic acid-based LFTs are limited in specificity and multiplexing capabilities, thus requiring improvements in accuracy and efficiency. In this study, we developed a dual lateral flow assay utilizing a thermophilic Argonaute, enabling the detection of multiple nucleic acids on a single strip with a sensitivity of 3 copies/μL and high specificity. Clinical validation showed 100 % concordance with commercial RT-qPCR results. This method not only preserves the high sensitivity of nucleic acid detection but also provides the benefits of portability, dual-target detection, and visual readout, highlighting its potential for point-of-care applications.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100321"},"PeriodicalIF":6.5,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143783162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Boundary engineering of porous zinc oxide nanosheets to enhance gas sensing performance 多孔氧化锌纳米片的边界工程增强气敏性能
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-04-02 DOI: 10.1016/j.snr.2025.100322
Min Young Kim , Haeun Choa , Chul Oh Park , Dung Thi Hanh To , Tran Khanh Phuong Cao , Emily Resendiz Mendoza , Changhyun Jin , Beom Zoo Lee , Nosang Vincent Myung , Kyu Hyoung Lee
{"title":"Boundary engineering of porous zinc oxide nanosheets to enhance gas sensing performance","authors":"Min Young Kim ,&nbsp;Haeun Choa ,&nbsp;Chul Oh Park ,&nbsp;Dung Thi Hanh To ,&nbsp;Tran Khanh Phuong Cao ,&nbsp;Emily Resendiz Mendoza ,&nbsp;Changhyun Jin ,&nbsp;Beom Zoo Lee ,&nbsp;Nosang Vincent Myung ,&nbsp;Kyu Hyoung Lee","doi":"10.1016/j.snr.2025.100322","DOIUrl":"10.1016/j.snr.2025.100322","url":null,"abstract":"<div><div>Zinc oxide (ZnO) based gas sensors are noted for their high sensing response toward a wide range of gases, however, they often face challenges in achieving other required properties due to complex reaction dynamics under different operating conditions. Herein, we fabricated boundary-engineered porous ZnO nanosheets by controlling calcination temperature of hydrozincite. These materials were systematically evaluated their sensing performance towards oxidizing gases, reducing inorganic gases, and volatile organic compounds. Additionally, detailed sensing properties toward NO<sub>2</sub>, NH<sub>3</sub>, and acetone were examined to investigate the critical roles of the energy barrier for carrier transport (<em>E</em><sub>b</sub>) and the nature of ionized oxygen species in the sensing materials. Characteristics of the materials, such as crystallite size, pore size, porosity, and surface area, were effectively tuned, which significantly influenced <em>E</em><sub>b</sub> and sensing performance. The porous ZnO nanosheets calcined at 300 °C for 4 h, exhibiting a higher density of boundaries, demonstrated enhanced sensing performance with maximum response values of 10.59 for NO<sub>2</sub>, 0.38 for NH<sub>3</sub>, and 0.41 for acetone at personnel exposure level. This improvement is likely linked to the increased surface charge carrier concentration and the higher <em>E</em><sub>b</sub> associated with greater boundary density, indicating an increase in available reaction active sites. Furthermore, our findings reveal that gas sensing behavior is strongly dependent on the type of ionized oxygen species present. The optimal sensing temperature for NO<sub>2</sub> is 250 °C, where the dominant ionized oxygen species is <em>O</em><sup>−</sup>, while for NH<sub>3</sub> and acetone, the optimal temperature was 450 °C, where O<sup>2−</sup> is the predominant species.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100322"},"PeriodicalIF":6.5,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143785234","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Facile patterning of microfluidic paper-based analytical devices (μPADs) by dispensing propylene glycol methyl ether acetate (PGMEA) 分配丙二醇甲基醚乙酸酯(PGMEA)的微流控纸基分析装置(μPADs)的简易图案化
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-04-02 DOI: 10.1016/j.snr.2025.100323
Xionghui Li , Xuanying Liang , Haonan Li , Jing Song , Kanghui Li , Muyang Zhang , Huiru Zhang , Zhuoting Han , Lok Ting Chu , Weijin Guo
{"title":"Facile patterning of microfluidic paper-based analytical devices (μPADs) by dispensing propylene glycol methyl ether acetate (PGMEA)","authors":"Xionghui Li ,&nbsp;Xuanying Liang ,&nbsp;Haonan Li ,&nbsp;Jing Song ,&nbsp;Kanghui Li ,&nbsp;Muyang Zhang ,&nbsp;Huiru Zhang ,&nbsp;Zhuoting Han ,&nbsp;Lok Ting Chu ,&nbsp;Weijin Guo","doi":"10.1016/j.snr.2025.100323","DOIUrl":"10.1016/j.snr.2025.100323","url":null,"abstract":"<div><div>Facile patterning of microfluidic paper-based analytical devices (μPADs) is highly intriguing for researchers using μPADs for point-of-care diagnostics. For the first time, this work introduces the use of propylene glycol methyl ether acetate (PGMEA) as a novel material to create barriers on μPADs, with the formation mechanism of these barriers investigated using scanning electron microscopy (SEM). A digitized and automated dispensing machine is employed for barrier creation, demonstrating the stability of the barriers in the presence of various organic and aqueous solvents including alkaline and acidic solutions, and surfactants. Additionally, we explore the possibility of rapid patterning of μPADs by hand drawing using PGMEA. The stability of the barriers created by PGMEA lays a solid foundation for various biomedical and chemical analyses. Moreover, we use this method to prepare μPADs for quantitative measurement of iodate content in saline solutions and blood typing analysis successfully, proving that μPADs prepared by this method have a significant potential for applications in point-of-care diagnostics.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100323"},"PeriodicalIF":6.5,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Meander-shaped piezoelectric MEMS loudspeaker with maximized area efficiency for in-ear applications 弯曲形状的压电MEMS扬声器,具有最大的面积效率,用于入耳应用
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-03-28 DOI: 10.1016/j.snr.2025.100319
Dennis Becker , Robert Scharf , Tobias Leonhard , Andreas Merz , Achim Bittner , Alfons Dehé
{"title":"Meander-shaped piezoelectric MEMS loudspeaker with maximized area efficiency for in-ear applications","authors":"Dennis Becker ,&nbsp;Robert Scharf ,&nbsp;Tobias Leonhard ,&nbsp;Andreas Merz ,&nbsp;Achim Bittner ,&nbsp;Alfons Dehé","doi":"10.1016/j.snr.2025.100319","DOIUrl":"10.1016/j.snr.2025.100319","url":null,"abstract":"<div><div>State-of-the-art MEMS loudspeakers use innovative actuator concepts to improve the acoustic performance from minimal chip volumes. Lateral vibration of multiple vertical actuators is a promising lever to increase the active emitting area while keeping the chip dimensions constant. Recent publications on this topic require a multiple wafer fabrication process to realize such actuator principles. This work demonstrates a new concept fabricating a closed folded membrane with vertical actuators in a single wafer process. Conformal deposition of piezoelectric aluminum nitride (AlN) thin films allow the lateral actuation of the transducer. Laser-doppler-vibrometer (LDV) measurements show that no actuator resonances are located below 40 kHz. Acoustic characterization of a manufactured device in an IEC ear simulator demonstrates a linear behavior of this loudspeaker with THD values below 1.5 % over the audible frequency range. At 1 kHz, a SPL of 64 dB and a THD of 0.3 % are measured with a driving voltage of 16 V<sub>p</sub>. A comparison of the acquired frequency response with simulated results shows a very good fit with deviations of less than 3.2 dB, successfully proving the concept and the potential of the folded MEMS loudspeaker.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100319"},"PeriodicalIF":6.5,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143738718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Nucleic acid-free aptamer-CRISPR/Cas14 biosensor for prosthetic joint infection rapid detection 无核酸适体crispr /Cas14生物传感器用于假体关节感染的快速检测
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-03-24 DOI: 10.1016/j.snr.2025.100318
Zong-Jie Geng , Quan-Bo Ji , Yan-Jing Chen , Lin Hao , Jun-Song Wang , Ze-Yu Feng , Qing-Yuan Zheng , Guo-Qiang Zhang , Yan Wang
{"title":"Nucleic acid-free aptamer-CRISPR/Cas14 biosensor for prosthetic joint infection rapid detection","authors":"Zong-Jie Geng ,&nbsp;Quan-Bo Ji ,&nbsp;Yan-Jing Chen ,&nbsp;Lin Hao ,&nbsp;Jun-Song Wang ,&nbsp;Ze-Yu Feng ,&nbsp;Qing-Yuan Zheng ,&nbsp;Guo-Qiang Zhang ,&nbsp;Yan Wang","doi":"10.1016/j.snr.2025.100318","DOIUrl":"10.1016/j.snr.2025.100318","url":null,"abstract":"<div><div>Rapid detection of live pathogens in prosthetic joint infections (PJI) remains a critical challenge in clinical practice. This study introduces a novel dual-aptamer CRISPR/Cas14 biosensor for rapid, equipment-minimized detection of live bacteria in clinical samples. Unlike existing aptamer-CRISPR platforms, our method integrates two aptamers: one biotinylated for magnetic bead-based bacterial enrichment, and the other to activate the CRISPR/Cas14 system. Cas14, chosen for its PAM-free ssDNA recognition and direct detection capability, triggers trans-cleavage of fluorophore-quencher-labeled ssDNA, generating fluorescence signals without nucleic acid extraction or amplification. This approach eliminates the need for centrifugation, significantly reducing equipment requirements and making it ideal for point-of-care testing. The method demonstrated high specificity, with detection limits of 10² CFU/mL for aptamer enrichment and 10 CFU/mL for CRISPR/Cas14 detection at room temperature (37 °C) . Clinical validation showed an 83.3 % positive concordance rate and 100 % negative concordance, with results obtainable within 15 min. By bypassing nucleic acid extraction and amplification, this dual-aptamer CRISPR/Cas14 biosensor offers a simple, rapid, and reliable solution for diagnosing PJI, with significant potential for clinical application.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100318"},"PeriodicalIF":6.5,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143725371","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Novel L-cysteine-functionalized Au@MnO2/MoO3 nanocomposites for electrochemical detection of apoptosis-related proteins in immunogenic cell death of patient-derived lung cancer cells 新型l -半胱氨酸功能化Au@MnO2/MoO3纳米复合材料用于电化学检测患者源性肺癌细胞免疫原性死亡中凋亡相关蛋白
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-03-19 DOI: 10.1016/j.snr.2025.100316
Yi-An Chen , Ming-You Shie , Kai-Bo Chang , Chao-Ming Su , Chia-Che Ho , Sheng-Wen Ye , Yi-Wen Chen
{"title":"Novel L-cysteine-functionalized Au@MnO2/MoO3 nanocomposites for electrochemical detection of apoptosis-related proteins in immunogenic cell death of patient-derived lung cancer cells","authors":"Yi-An Chen ,&nbsp;Ming-You Shie ,&nbsp;Kai-Bo Chang ,&nbsp;Chao-Ming Su ,&nbsp;Chia-Che Ho ,&nbsp;Sheng-Wen Ye ,&nbsp;Yi-Wen Chen","doi":"10.1016/j.snr.2025.100316","DOIUrl":"10.1016/j.snr.2025.100316","url":null,"abstract":"<div><div>This study presents the development of a label-free electrochemical immunosensor for detecting apoptosis-related proteins, calreticulin (CRT) and high-mobility group box 1 (HMGB1), in the context of immunogenic cell death (ICD). The immunosensor utilizes a novel <em>L</em>-Cys/Au@MnO<sub>2</sub>/MoO<sub>3</sub> nanocomposite-modified screen-printed carbon electrode (SPCE) to investigate the electrochemical detection of HMGB1, a topic with limited prior research. The successful assembly of the immunosensor was confirmed through XRD, SEM, and XPS analyses. The strong biomolecular adsorption capability of Au nanoparticles, combined with the superior charge transfer properties of MnO<sub>2</sub> and MoO<sub>3</sub>, enhances the electrochemical performance of the immunosensor. The immunosensor achieved low detection limits (<em>LOD</em>) of 0.5391 pg/mL for CRT and 0.849 pg/mL for HMGB1, along with broad dynamic linear ranges of 0.001 to 10 ng/mL and 0.001 to 1000 ng/mL, respectively, highlighting the excellent selectivity, reproducibility, and stability of this novel immunosensor. With higher drug concentrations and extended treatment durations, increased levels of ICD biomarkers were observed, underscoring the potential of the immunosensor for monitoring drug efficacy and its applicability in personalized medicine.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100316"},"PeriodicalIF":6.5,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143705905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A BRET-based Mpro biosensor containing a nanobody and tandem cleavage sites shows an increased cleavage rate 含有纳米体和串联切割位点的基于bret的Mpro生物传感器显示出更高的切割速率
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-03-15 DOI: 10.1016/j.snr.2025.100315
Anupriya M Geethakumari , Asfia Sultana , Asma Fatima , S M Nasir Uddin , Somaiya Abdulhakim , Amera Mohamed , Samiha Rahman , Khaloud Al-Buainain , Hadi M Yassine , Hebah A Al Khatib , Kabir H Biswas
{"title":"A BRET-based Mpro biosensor containing a nanobody and tandem cleavage sites shows an increased cleavage rate","authors":"Anupriya M Geethakumari ,&nbsp;Asfia Sultana ,&nbsp;Asma Fatima ,&nbsp;S M Nasir Uddin ,&nbsp;Somaiya Abdulhakim ,&nbsp;Amera Mohamed ,&nbsp;Samiha Rahman ,&nbsp;Khaloud Al-Buainain ,&nbsp;Hadi M Yassine ,&nbsp;Hebah A Al Khatib ,&nbsp;Kabir H Biswas","doi":"10.1016/j.snr.2025.100315","DOIUrl":"10.1016/j.snr.2025.100315","url":null,"abstract":"<div><div>Here, we report the engineering of a Bioluminescence Resonance Energy Transfer (BRET)-based SARS-CoV-2 main protease (M<sup>pro</sup>) biosensor containing the M<sup>pro</sup> N-terminal autocleavage sequence in tandem and a nanobody that shows an enhanced rate of M<sup>pro</sup>-mediated proteolytic cleavage. Specifically, we designed M<sup>pro</sup> biosensors containing 2×, 4× and 8× repeats of M<sup>pro</sup> N-terminal autocleavage sequences and a combination of M<sup>pro</sup> cleavage sequences containing a total of 12 cleavage sites sandwiched between mNeonGreen (mNG) and NanoLuc (NLuc). Gaussian accelerated molecular dynamics (GaMD) simulations of the predicted alpha-helical synthetic M<sup>pro</sup> cleavage sequences revealed a dynamic nature of the cleavage sequences, which is critical for their efficient cleavage, and a relatively short end-to-end distances, which is required for high BRET. Live cell assays revealed a cleavage sequence length-dependent resonance energy transfer, except for the 12× -syn cleavage site, and an increased rate of cleavage and a decreased pharmacological inhibitor efficacy for the M<sup>pro</sup> biosensor containing 2× cleavage sequences. Further, mutational analysis revealed a requirement for both cleavage sites to be intact for increased cleavage rate. Importantly, the inclusion of an M<sup>pro</sup>-binding, but non-inhibiting, NB2E3 nanobody at the N-terminal further increased the cleavage rate of the 2× cleavage sequence-containing M<sup>pro</sup> biosensor. We envisage that the NB2E3 nanobody-2× M<sup>pro</sup> biosensor engineered here will be useful in drug discovery and functional characterization of M<sup>pro</sup> mutants in newly emerging SARS-CoV-2 variants as well as in detecting SARS-CoV-2 infection in a point-of-care testing (POCT) format.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100315"},"PeriodicalIF":6.5,"publicationDate":"2025-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143684185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
DNA nanotechnology and catalytic hairpin assembly cascade: A novel approach in achieving sensitive detection and in situ imaging of phosphatidylserine on living cell membranes DNA纳米技术和催化发夹组装级联:实现活细胞膜上磷脂酰丝氨酸的敏感检测和原位成像的新方法
IF 6.5
Sensors and Actuators Reports Pub Date : 2025-03-13 DOI: 10.1016/j.snr.2025.100314
Yiwei Ying , Wenxin Qi , Zixin Chen , Junyi Zhuang , Chang Feng , Robert Chunhua Zhao , Jiao Wang
{"title":"DNA nanotechnology and catalytic hairpin assembly cascade: A novel approach in achieving sensitive detection and in situ imaging of phosphatidylserine on living cell membranes","authors":"Yiwei Ying ,&nbsp;Wenxin Qi ,&nbsp;Zixin Chen ,&nbsp;Junyi Zhuang ,&nbsp;Chang Feng ,&nbsp;Robert Chunhua Zhao ,&nbsp;Jiao Wang","doi":"10.1016/j.snr.2025.100314","DOIUrl":"10.1016/j.snr.2025.100314","url":null,"abstract":"<div><div>Phosphatidylserine (PS) is a vital lipid in eukaryotic cell membranes, and its content variations are closely associated with various diseases. Therefore, tracking PS in living cells is invaluable. Here, we designed an aptamer-initiated cascade catalytic hairpin assembly (CHA)-based DNA triangular prism (AI-CDTP) for sensitive detection and in situ imaging of PS on living cell membranes. The aptamer-initiated DNA triangular prism (AI-DTP) was prepared as precursor probe using several single-stranded DNA through a simple one-pot annealing process, which takes only a few minutes. Additionally, the sensitivity of the PS-triggered CHA circuit was enhanced by coupling the hairpin probes on AI-DTP. This coupling increased the local concentration and accelerated the reaction kinetics, achieving a detection limit as low as 1.39 nM. Compared with the traditional plasmid systems, AI-CDTP simplifies the preparation process, responds rapidly, and demonstrates excellent self-delivery, biocompatibility and stability. Thus, our developed strategy provides a reliable platform for the sensitive imaging of intracellular PS.</div></div>","PeriodicalId":426,"journal":{"name":"Sensors and Actuators Reports","volume":"9 ","pages":"Article 100314"},"PeriodicalIF":6.5,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143684183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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