Biosensors and Bioelectronics最新文献

{"title":"\"Turn-on\" aptamer-immune lateral flow assays for the detection of small molecule targets based on CHA-assisted and CRISPR/Cas12a mediated signal transduction and amplification","authors":"Haowei Dong ,&nbsp;Yifei Qin ,&nbsp;Pengwei Zhang ,&nbsp;Liwen Lv ,&nbsp;Chunlei Yu ,&nbsp;Peng Jia ,&nbsp;Jicheng Zhao ,&nbsp;Fangling Du ,&nbsp;Yemin Guo ,&nbsp;Xia Sun","doi":"10.1016/j.bios.2025.117593","DOIUrl":"10.1016/j.bios.2025.117593","url":null,"abstract":"<div><div>Lateral flow assays (LFAs) have emerged as crucial tools for on-site food safety detection due to their simple operation and intuitive detection results. Nevertheless, LFAs for small molecule targets such as pesticides often present a \"Turn-off\" signal output, which leads to their low sensitivity and the risk of false positives. In this study, a CRISPR/Cas12a system-mediated strategy was employed to convert aptamer signals into the signals of immune LFAs, achieving a \"Turn-on\" signal output for highly sensitive detection of small molecule targets. The binding of aptamers to targets released the trigger sequence to initiate the catalytic hairpin assembly (CHA) reaction, generating double-stranded DNA, which subsequently activated the CRISPR/Cas12a system to cleave the FAM-labeled Reporter. Eventually, the \"Turn-on\" visual output of the signal was realized through an anti-6-FAM immune LFAs. The experiment optimized the sample pool preparation, CHA reaction conditions, CRISPR/Cas12a activation parameters, and the assembly process of the LFAs. The limit of detection for procymidone was as low as 0.015 ng/mL, which was 52.67 times more sensitive than those of conventional aptamer-based LFAs without signal amplification strategies. This method exhibits high specificity for procymidone and a recovery rate ranging from 94.00% to 104.20% in vegetable samples, demonstrating excellent stability and practicability.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"285 ","pages":"Article 117593"},"PeriodicalIF":10.7,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144106734","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}

{"title":"Enhanced electrochemical activity by MOF superstructure derived Ni2P@C for ultrasensitive sensing of Bisphenol A","authors":"Pan Gao ,&nbsp;Mian Zahid Hussain ,&nbsp;David Gryc ,&nbsp;Soumya Mukherjee ,&nbsp;Zhenyu Zhou ,&nbsp;Weijin Li ,&nbsp;Andreas Jentys ,&nbsp;Martin Elsner ,&nbsp;Roland A. Fischer","doi":"10.1016/j.bios.2025.117598","DOIUrl":"10.1016/j.bios.2025.117598","url":null,"abstract":"<div><div>Electrochemical (EC) sensing of bisphenol A (BPA), a notorious persistent contaminant, is of pressing interest. However, the state-of-the-art BPA sensors are challenged by two performance parameters: limited EC catalysis and sensitivity. Herein, a two-dimensional (2D) metal-organic framework (MOF) superstructure-derived Ni₂P@C probe elicits a novel EC sensor that exhibits high-efficiency BPA detection. Thanks to the abundant Ni^δ+ active sites exposed uniformly on cross-linked layers stemming from the inherited 2D-MOF superstructures as the precursors, high conductivity results from the organic linkers-derived graphitic carbon. The prepared Ni₂P@C composites-based EC sensors demonstrated exceptional BPA-induced sensing responses with a wide dynamic response range, high sensitivity of 0.951 μA cm⁻²·<em>μ</em>M⁻¹, a low limit of detection (LOD, 56.8 nM) in the linear range of 1 <em>μ</em>M–100 μM. Below 1 μM, the response followed the logarithm of BPA concentrations, indicating the potential for detection down to 5 pM. The excellent selectivity in the presence of similar interferents, combined with high reproducibility and chemical stability, underscores the potential of 2D MOF-derived Ni₂P@C for accurate monitoring of hazardous phenols, opening new avenues for environmental sensing and remediation.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"286 ","pages":"Article 117598"},"PeriodicalIF":10.7,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144147135","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}

{"title":"CapSense-MIP: Self-operating molecularly imprinted polymer (MIP) biosensor for point-of-care diagnostics","authors":"Azam Zare ,&nbsp;Bahareh Babamiri ,&nbsp;Mohsen Hassani ,&nbsp;Mahmood Khalghollah ,&nbsp;Mehdi Mohammadi ,&nbsp;Shaghayegh Haghjooy Javanmard ,&nbsp;Amir Sanati Nezhad","doi":"10.1016/j.bios.2025.117599","DOIUrl":"10.1016/j.bios.2025.117599","url":null,"abstract":"<div><div>Molecularly imprinted polymer (MIP) biosensors hold immense promise for point-of-care (POC) diagnostics due to their exceptional robustness, long shelf-life stability, selectivity, and ability to detect diverse biomarkers across their (patho)physiological ranges. However, their full potential within practical POC devices remains constrained by technical challenges, including the need for precise incubation control, effective washing of non-specific bindings, and consistent fluid handling in miniaturized systems. Without addressing these limitations, their ability to reliably operate in complex bodily fluids and within critical physiological detection ranges is significantly compromised. To address these barriers, we developed the CapSense-MIP, a capillary microfluidic biosensing platform that automates key MIP biosensing steps—sample aliquoting, incubation, washing, and sensing—using a fully integrated and self-powered system driven by sequential liquid delivery to the biosensing chamber. It incorporates an inlet regulator for precise aliquoting and incubation control, a suction-enhanced fiber for optimized washing, and the regulated channels and valve network to ensure high sensitivity and reproducibility, and a wide dynamic detection range that meets (patho)physiological needs. As a proof of concept, the CapSense-MIP was validated for agmatine detection, achieving a wide linear range of 1.0 nM–10 μM and an impressive limit of detection of 0.1 nM, with optimized protocols for both phosphate-buffered saline and human plasma. By addressing these critical challenges and leveraging the long shelf-life stability of MIP biosensors, the CapSense-MIP not only eliminates traditionally manual workflows but also delivers scalable, rapid, and cost-effective solutions for POC diagnostics, unlocking transformative potential for applications in healthcare and environmental monitoring.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"286 ","pages":"Article 117599"},"PeriodicalIF":10.7,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144167180","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}

{"title":"Engineered Cas12a-based one-tube detection of DNMT3A R882 H/C mutation in acute myeloid leukemia","authors":"Yue Liu ,&nbsp;Yin Liu ,&nbsp;Sanyun Wu ,&nbsp;Rui Cao ,&nbsp;Yunbao Pan ,&nbsp;Fuling Zhou","doi":"10.1016/j.bios.2025.117609","DOIUrl":"10.1016/j.bios.2025.117609","url":null,"abstract":"<div><div>Advances in sequencing technologies have identified numerous genetic alterations associated with acute myeloid leukemia (AML), many of which play critical roles in diagnosis, classification, and prognosis. Among these, mutations in the DNA methyltransferase 3 alpha (<em>DNMT3A</em>) gene are particularly prevalent, with the R882H and R882C variants being the most common. Accurate and sensitive detection of <em>DNMT3A</em> mutations is crucial for prognosis, treatment guidance, and early intervention in AML. However, existing detection methods often fail to achieve an optimal balance among sensitivity, turnaround time, and operational simplicity. To address this limitation, we aimed to develop a rapid and highly sensitive method for detecting <em>DNMT3A</em> mutations. The CRISPR/Cas12a system shows promise for genetic detection due to its high sensitivity and single-base specificity. Here, we established a Cas12a-based one-tube assay for the detection of <em>DNMT3A</em> R882 H/C mutations. We utilized the mismatch tolerance of enAsU-R Cas12a to design crRNA for DNMT3A R882 H/C mutation and integrated CRISPR/Cas12a system with ERA. The entire detection process can be completed within 1 h at 37 °C. The optimized detection system demonstrated a sensitivity of 0.1 % when analyzing genomic DNA. To validate its clinical applicability, we tested samples from 49 AML patients and successfully identified all <em>DNMT3A</em> R882H/C-positive cases, including one with a mutation rate as low as 0.24 %. These results highlight the potential of our Cas12a-based one-tube detection system as a rapid, sensitive, and cost-effective method for detecting <em>DNMT3A</em> R882 H/C mutation. This approach could serve as a valuable tool for both diagnostic and therapeutic monitoring.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"286 ","pages":"Article 117609"},"PeriodicalIF":10.7,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124901","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}

{"title":"Changing complexity into simplicity: visualization of endoplasmic reticulum autophagy employing a self-deliver viscosity sensor by lifetime imaging","authors":"Nan Sheng ,&nbsp;Jie Niu ,&nbsp;Xingyu Lyu ,&nbsp;Yangang Su ,&nbsp;Qing Lyu ,&nbsp;Wendong Jin ,&nbsp;Miao Cheng ,&nbsp;Jiaxuan Wang ,&nbsp;Li Li ,&nbsp;Xiaoqiang Yu","doi":"10.1016/j.bios.2025.117596","DOIUrl":"10.1016/j.bios.2025.117596","url":null,"abstract":"<div><div>Endoplasmic reticulum autophagy (ER-phagy) is a highly specialized form of autophagy that maintains cellular homeostasis by sweeping damaged or excessive endoplasmic reticulum segments. Dysregulation of ER-phagy has profound implications for normal physiological activities and is closely associated with various diseases. Therefore, accurately visualizing ER-phagy process is imperative. Herein, we successfully developed a novel ER-targeted viscosity sensor (<strong>YKI</strong>) for conveniently and accurately visualizing ER-phagy by lifetime imaging. Compared with commercial ER Tracker Green and ER Tracker Red that required specific combination incubation reagent to stain the endoplasmic reticulum of live cell, <strong>YKI</strong> could directly target endoplasmic reticulum by the self-delivery way and without leakage, which profited from the hydrophobic interaction between the long alkyl chain of <strong>YKI</strong> and the lipid bilayer of the ER. And it could also specifically target the endoplasmic reticulum of fixed cells. Since <strong>YKI</strong> was sensitive to viscosity, it displayed short fluorescence lifetime in endoplasmic reticulum but exhibited longer fluorescence lifetime upon co-delivery with ER into lysosomes. As a consequence, <strong>YKI</strong> could achieve accurate and convenient visualization of ER-phagy alone <em>via</em> lifetime imaging. This work provided a powerful tool for studying ER-phagy and could promote the development of related fields.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"286 ","pages":"Article 117596"},"PeriodicalIF":10.7,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144116590","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}

{"title":"Micro-scale thermofluidics enable autonomous and scalable CRISPR diagnostics for sexually transmitted infections screening","authors":"Ling Lin ,&nbsp;Yaohua Xue ,&nbsp;Lufeng Tan ,&nbsp;Cheng Jiang ,&nbsp;Mingxu Liu ,&nbsp;Xinying Li ,&nbsp;Jieyu Qiu ,&nbsp;Huizhen Zhang ,&nbsp;Jiajian Zhou ,&nbsp;Bowen Shu","doi":"10.1016/j.bios.2025.117591","DOIUrl":"10.1016/j.bios.2025.117591","url":null,"abstract":"<div><div>The development of clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid detection has recently been a center of interest for next-generation molecular diagnostics. Despite considerable advances, simple and effective strategies to harness the isothermal amplification reaction and CRISPR-based detection for maximal performance and minimal complexity are still desirable. Here, a thermofluidic approach leverages the micro-scale chemical and physical mechanism to perform autonomous and scalable CRISPR-based diagnostics (CRISPR-Dx) in a greatly simplified format, which was called “Thermofluidic CRISPR”. Originating from the concept of convective PCR, it utilizes looped microchannel reactors to perform approximatively undisturbed isothermal amplification reaction at balanced temperature by virtue of the restricted molecular diffusion across the microchannel, in which the reagents of two reactions are compartmentalized virtually; then it creates circulatory flow within the loop channel to mix the amplificons and CRISPR reagents <em>via</em> Rayleigh–Bénard thermal convection, by simply warming up one side of the loop channel. Due to the simplicity and scalability, a low-cost, battery-powered, portable diagnostic platform, incorporating with smartphone-enabled real-time fluorescence readout, to perform rapid (&lt;30 min), highly sensitive (2 copies per reaction), quantitative and multiplexed CRISPR-Dx was constructed. Its diagnostic performance in rapid screening of multiple pathogens from 196 clinical samples for syndromic testing of sexually transmitted infections was evaluated, exhibiting 97.4 % sensitivity and 100 % specificity benchmarked against the laboratory-based testing. Leveraging the micro-scale chemical and physical mechanism to simplify workflows for CRISPR-Dx may enhance their versatility and facilitate their broader applicability at the point of care.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"285 ","pages":"Article 117591"},"PeriodicalIF":10.7,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144107090","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}

{"title":"An innovative electrophoresis-coupled electrochemiluminescence immunosensor for rapid and sensitive detection of carcinoembryonic antigen","authors":"Xiaoting Wu ,&nbsp;Qing Lu ,&nbsp;Shu Zhu ,&nbsp;Shengnan Tang ,&nbsp;Yusha Li ,&nbsp;Lei Ma ,&nbsp;Xiaoqing Ming ,&nbsp;Wei Jiang ,&nbsp;Zhanghong Wu ,&nbsp;Jinying Hu ,&nbsp;Xiaorui Huang ,&nbsp;Jing Huang ,&nbsp;Jianjun Hu ,&nbsp;Yuchan Zhang ,&nbsp;Guangchao Zang","doi":"10.1016/j.bios.2025.117595","DOIUrl":"10.1016/j.bios.2025.117595","url":null,"abstract":"<div><div>Electrochemiluminescence (ECL) immunosensor provides unique advantages for the sensitive biomarker detection. However, the lengthy detection duration and plentiful system interferences have hindered their development. Here, we present an innovative approach in which electrophoresis coupled with ECL immunoassay was implemented to construct an electric field-enhanced ECL immunosensor for efficient detection of carcinoembryonic antigen (CEA). The electrophoresis device can be regarded as an electric field-driven incubation system, with a working electrode as the anode and a platinum disk as the cathode. Upon applying direct voltage, CEA was swiftly transported to the electrode surface via an upward electric field force, drastically cutting the CEA incubation time from 60 min to just 5 min—a 12-fold reduction compared to traditional methods. Our method also achieved a broad linear detection range from 10⁻² to 10⁴ pg/mL, with a lower detection limit of 2.33 fg/mL. Additionally, we utilized the COMSOL Multiphysics-based numerical model and substantial experiment results, demonstrating that the incorporation of an electrophoresis system has allowed for rapid detection with high sensitivity, thereby boosting the overall efficiency of the ECL immunosensor. This study underscores the potential of the electric field-enhanced ECL immunosensor for broad application in the biodetection field.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"286 ","pages":"Article 117595"},"PeriodicalIF":10.7,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144131033","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}

{"title":"Stellate silicon microneedles for rapid point-of-care melanoma exosome isolation and detection via a lateral flow assay","authors":"Scott M. Smith ,&nbsp;Abha Kumari ,&nbsp;Joseph P. Marvar,&nbsp;Nna-Emeka Onukwugha,&nbsp;Yoon-Tae Kang,&nbsp;Sunitha Nagrath","doi":"10.1016/j.bios.2025.117560","DOIUrl":"10.1016/j.bios.2025.117560","url":null,"abstract":"<div><div>Melanoma is the most aggressive type of skin cancer with high mortality rates. Early diagnosis is crucial because it significantly improves treatment outcomes, but conventional methods relying on dermoscopy and lesion biopsy have limitations in accuracy during early stages and are invasive. Liquid biopsies offer a minimally invasive alternative, particularly for routine screening. The abundance of cancer cell-driven extracellular vesicles in interstitial fluid can be utilized for point-of-care cancer diagnostics. Here, we developed a stellate silicon microneedle patch, the ExoPatch, coated with Annexin V functionalized hydrogel to isolate melanoma-specific exosomes. The ExoPatch captures exosomes directly from the skin, followed by dissolution of the hydrogel to release the exosomes, which are then detected using a lateral flow immunoassay specific to melanoma markers (MCAM and MCSP). After validating with cell line derived extracellular vesicles and testing with mouse tissue, the ExoPatch isolated 11.5 times more protein from melanoma tissue compared to healthy tissue. Additionally, the ExoPatch effectively distinguished between melanoma and healthy tissues, with its specificity confirmed through Western Blot and electron microscopy analysis. The ExoPatch with melanoma mouse samples produced a 3.5-fold higher signal in the lateral flow immunoassay compared to that of healthy controls. The ExoPatch presents a promising point-of-care diagnostic tool for melanoma, offering significant advantages in terms of rapidness, minimal invasiveness, and ease of use. It has the potential to enhance early detection and routine monitoring in melanoma patients, ultimately improving patient outcomes by reducing the reliance on traditional, invasive biopsies.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"285 ","pages":"Article 117560"},"PeriodicalIF":10.7,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144106733","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}

{"title":"Advanced droplet microfluidic platform for high-throughput screening of industrial fungi","authors":"Qiaoyi Yang ,&nbsp;Siqi Lu ,&nbsp;Haoyu Wu ,&nbsp;Danshan Zhao ,&nbsp;Wei Wei ,&nbsp;Haoran Yin ,&nbsp;Xiang Li ,&nbsp;Chao Ye ,&nbsp;Tianqiong Shi ,&nbsp;Zhe Wang ,&nbsp;Yuetong Wang","doi":"10.1016/j.bios.2025.117594","DOIUrl":"10.1016/j.bios.2025.117594","url":null,"abstract":"<div><div>Industrial fungi are pivotal candidates for the production of a diverse array of bioproducts. To enhance their productivity, these strains are frequently subjected to genetic modifications. Following transformation, the selection of optimal production strains is critical; however, traditional screening methods often suffer from limitations in throughput and sensitivity. This article explores the transformative potential of Droplet Microfluidic Technology (DMFS) for high-throughput screening of industrial fungi. DMFS enables real-time monitoring and precise single-cell analysis by encapsulating individual fungal spores or cells within droplets, ranging from picoliters to nanoliters, functioning as isolated microreactors. This technology effectively addresses the challenges posed by conventional methods, such as agar plate assays and fluorescence-activated cell sorting. Key advancements discussed include microfluidic chip fabrication, droplet generation and regulation techniques, and multimodal signal detection methods—encompassing fluorescence, Raman spectroscopy, and mass spectrometry. Notably, strategies to mitigate droplet breakage in filamentous fungi, including physical constraints, bionic core-shell hydrogels, and genetic engineering approaches, are analyzed to prolong stable culture times. Future developments will likely emphasize interdisciplinary applications, including automation driven by artificial intelligence and label-free detection methods. We anticipate that this review will catalyze further research into high-quality industrial fungi, thereby promoting sustainable biomanufacturing through enhanced throughput, cost-effectiveness, and scalability.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"285 ","pages":"Article 117594"},"PeriodicalIF":10.7,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144106732","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}

{"title":"Peptide-based biosensor for real-time monitoring of protease biomarker activity using multi-parametric surface plasmon resonance spectroscopy","authors":"Pratika Rai ,&nbsp;Sabrina N. Hoba ,&nbsp;Celine Buchmann ,&nbsp;Christian Kersten ,&nbsp;Tanja Schirmeister ,&nbsp;Bernd Bufe ,&nbsp;Alexey Tarasov","doi":"10.1016/j.bios.2025.117586","DOIUrl":"10.1016/j.bios.2025.117586","url":null,"abstract":"<div><div>Matrix metalloproteinase-9 (MMP-9) is a key biomarker targeted in biosensing applications due to its involvement not only in maintaining good health but also in triggering various diseases such as cancer. While quantitative detection of MMP-9 is widely performed using bioanalytical detection kits such as enzyme-linked immunosorbent assay (ELISA), faster, label-free and real-time monitoring of MMP-9 activity would lead to improved disease diagnosis with better understanding of its role in underlying disease progression and development of therapeutic strategies. In this work, multi-parametric surface plasmon resonance spectroscopy (MP-SPR) is used to develop a highly sensitive MMP-9 sensor using immobilized synthetic peptides as MMP-9 substrates. Upon binding to MMP-9, the MMP-9 specific peptide is hydrolyzed between two sites of the amino acid sequence (P1 Gly and P1′ Met), resulting in a decrease in the SPR signal response. The sensor detects different concentrations of MMP-9 in buffer and cell culture medium (RPMI-1640), indicating that it can be used under physiological conditions. The limit of detection (LOD) for MMP-9 in buffer is 0.34 pM and the linear detection range is between 5 pM and 9 nM, covering the clinically relevant detection range of MMP-9. To our knowledge, this is the first short synthetic peptide-based MP-SPR biosensor for monitoring MMP-9 activity. The sensor is faster than ELISA (minutes vs. hours) and provides real-time detection with access to binding kinetics information. The use of MP-SPR provides information on surface coverage and peptide thickness before and after cleavage, which is unique compared to other detection methods.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"286 ","pages":"Article 117586"},"PeriodicalIF":10.7,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144135205","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}