Surface Plasmon Resonance (SPR)-based biodetection systems have emerged as powerful tools for real-time, label-free biomolecular interaction analysis, revolutionizing fields such as diagnostics, drug discovery, and environmental monitoring. This review highlights the foundational principles of SPR, focusing on the interplay of evanescent waves and surface plasmons that underpin its high sensitivity and specificity. Recent advancements in SPR technology, including enhancements in sensor chip materials, integration with nanostructures, and coupling with complementary detection techniques, are discussed to showcase their role in improving analytical performance. The paper also explores diverse applications of SPR biodetection systems, ranging from pathogen detection and cancer biomarker identification to food safety monitoring and environmental toxin analysis. By providing a comprehensive overview of technological progress and emerging trends, this review underscores the transformative potential of SPR-based biodetection systems in addressing critical scientific and societal challenges. Future directions and challenges, including miniaturization, cost reduction, and expanding multiplexing capabilities, are also presented to guide ongoing research and development in this rapidly evolving field.
{"title":"Surface Plasmon Resonance-Based Biodetection Systems: Principles, Progress and Applications-A Comprehensive Review.","authors":"Muhammad A Butt","doi":"10.3390/bios15010035","DOIUrl":"10.3390/bios15010035","url":null,"abstract":"<p><p>Surface Plasmon Resonance (SPR)-based biodetection systems have emerged as powerful tools for real-time, label-free biomolecular interaction analysis, revolutionizing fields such as diagnostics, drug discovery, and environmental monitoring. This review highlights the foundational principles of SPR, focusing on the interplay of evanescent waves and surface plasmons that underpin its high sensitivity and specificity. Recent advancements in SPR technology, including enhancements in sensor chip materials, integration with nanostructures, and coupling with complementary detection techniques, are discussed to showcase their role in improving analytical performance. The paper also explores diverse applications of SPR biodetection systems, ranging from pathogen detection and cancer biomarker identification to food safety monitoring and environmental toxin analysis. By providing a comprehensive overview of technological progress and emerging trends, this review underscores the transformative potential of SPR-based biodetection systems in addressing critical scientific and societal challenges. Future directions and challenges, including miniaturization, cost reduction, and expanding multiplexing capabilities, are also presented to guide ongoing research and development in this rapidly evolving field.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763797/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034671","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhuolun Meng, Hassan Raji, Mahtab Kokabi, Deng Zou, James Chan, Qihao Liu, Ruifeng Zhang, Mehdi Javanmard
CD4 T lymphocytes play a key role in initiating the adaptive immune response, releasing cytokines that mediate numerous signal transduction pathways across the immune system. Therefore, CD4 T cell counts are widely used as an indicator of overall immunological health. HIV, one of the leading causes of death in the developing world, specifically targets and gradually depletes CD4 cells, making CD4 counts a critical metric for monitoring disease progression. As a result, accurately counting CD4 cells represents a pressing challenge in global healthcare. Flow cytometry remains the gold standard for enumerating CD4 T cells; however, flow cytometers are expensive, difficult to transport, and require skilled medical staff to prepare samples, operate the equipment, and interpret results. This highlights the critical need for novel, rapid, cost-effective, and portable methods of CD4 enumeration that are suitable for deployment in resource-limited countries. This review will survey and analyze emerging research in CD4 counting, with a focus on microfluidic systems, which represent a promising area of investigation.
{"title":"Microfluidic Assays for CD4 T Lymphocyte Counting: A Review.","authors":"Zhuolun Meng, Hassan Raji, Mahtab Kokabi, Deng Zou, James Chan, Qihao Liu, Ruifeng Zhang, Mehdi Javanmard","doi":"10.3390/bios15010033","DOIUrl":"10.3390/bios15010033","url":null,"abstract":"<p><p>CD4 T lymphocytes play a key role in initiating the adaptive immune response, releasing cytokines that mediate numerous signal transduction pathways across the immune system. Therefore, CD4 T cell counts are widely used as an indicator of overall immunological health. HIV, one of the leading causes of death in the developing world, specifically targets and gradually depletes CD4 cells, making CD4 counts a critical metric for monitoring disease progression. As a result, accurately counting CD4 cells represents a pressing challenge in global healthcare. Flow cytometry remains the gold standard for enumerating CD4 T cells; however, flow cytometers are expensive, difficult to transport, and require skilled medical staff to prepare samples, operate the equipment, and interpret results. This highlights the critical need for novel, rapid, cost-effective, and portable methods of CD4 enumeration that are suitable for deployment in resource-limited countries. This review will survey and analyze emerging research in CD4 counting, with a focus on microfluidic systems, which represent a promising area of investigation.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763704/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Liliya I Mukhametova, Marya K Kolokolova, Ivan A Shevchenko, Boris S Tupertsev, Anatoly V Zherdev, Chuanlai Xu, Sergei A Eremin
2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the popular herbicides that is widely used in agriculture and can be found in food and water. A rapid and sensitive fluorescence polarization immunoassay (FPIA) was proposed for the detection of 2,4-D in juice and water. New tracers, 2,4-D-buthylenediamin fluoresceinthiocarbamyl (2,4-D-BDF) and 2,4-D-glycine aminofluorescein (2,4-D-GAF), were obtained and characterized. Monoclonal antibodies (MAb) obtained against 2,4-D were used as a recognition reagent. The kinetics of the interaction of MAb and tracers were studied, and the kinetic parameters of their binding were calculated. High specificity of binding of tracers and MAb was shown. In this work, an approach was elaborated on to reduce the detection limit of 2,4-D by the FPIA method by changing the volume of the studied sample. The optimized FPIA in a competitive format was characterized by the LODs of 2,4-D 8 and 0.4 ng/mL and the working ranges 30-3000 ng/mL and 3-300 ng/mL for juice and water, respectively. The entire test cycle (from sample receipt to evaluation of the analysis results) took only 20 min. The test for the recovery of 2,4-D in juice and water gave values from 95 to 120%, which demonstrated the reliability of the herbicide determination in real samples.
{"title":"Fluorescence Polarization Immunoassay for Rapid, Sensitive Detection of the Herbicide 2,4-Dichlorophenoxyacetic Acid in Juice and Water Samples.","authors":"Liliya I Mukhametova, Marya K Kolokolova, Ivan A Shevchenko, Boris S Tupertsev, Anatoly V Zherdev, Chuanlai Xu, Sergei A Eremin","doi":"10.3390/bios15010032","DOIUrl":"10.3390/bios15010032","url":null,"abstract":"<p><p>2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the popular herbicides that is widely used in agriculture and can be found in food and water. A rapid and sensitive fluorescence polarization immunoassay (FPIA) was proposed for the detection of 2,4-D in juice and water. New tracers, 2,4-D-buthylenediamin fluoresceinthiocarbamyl (2,4-D-BDF) and 2,4-D-glycine aminofluorescein (2,4-D-GAF), were obtained and characterized. Monoclonal antibodies (MAb) obtained against 2,4-D were used as a recognition reagent. The kinetics of the interaction of MAb and tracers were studied, and the kinetic parameters of their binding were calculated. High specificity of binding of tracers and MAb was shown. In this work, an approach was elaborated on to reduce the detection limit of 2,4-D by the FPIA method by changing the volume of the studied sample. The optimized FPIA in a competitive format was characterized by the LODs of 2,4-D 8 and 0.4 ng/mL and the working ranges 30-3000 ng/mL and 3-300 ng/mL for juice and water, respectively. The entire test cycle (from sample receipt to evaluation of the analysis results) took only 20 min. The test for the recovery of 2,4-D in juice and water gave values from 95 to 120%, which demonstrated the reliability of the herbicide determination in real samples.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763590/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034635","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Luciana de Souza Freire, Ariamna María Dip Gandarilla, Yonny Romaguera Barcelay, Camila Macena Ruzo, Barbara Batista Salgado, Ana P M Tavares, Francisco Xavier Nobre, Julio Nino de Souza Neto, Spartaco Astolfi-Filho, Ștefan Țălu, Pritesh Lalwani, Niranjan Patra, Walter Ricardo Brito
Biosensors harness biological materials as receptors linked to transducers, enabling the capture and transformation of primary biorecognition signals into measurable outputs. This study presents a novel carboxylation method for synthesizing carboxylated graphene (CG) under acidic conditions, enhancing biosensing capabilities. The characterization of the CG was performed using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Raman spectroscopy, thermogravimetric analysis (TGA), and X-ray diffraction (XRD). We modified screen-printed carbon electrodes (SPCEs) with CG to immobilize the SARS-CoV-2 N-protein, facilitating targeted detection of IgA antibodies (IgA-SARS-CoV-2). The analytical performance was assessed via electrochemical techniques such as cyclic voltammetry and electrochemical impedance spectroscopy, confirming CG synthesis effectiveness and biosensor functionality. The developed biosensor efficiently detects IgA-SARS-CoV-2 across a dilution range of 1:1000 to 1:200 v/v in a phosphate-buffered saline (PBS) solution, with a limit of detection calculated at 1:1601 v/v. This device shows considerable potential because of its fast response time, miniaturized design facilitated by SPCEs, reduced sample volume requirements, high sensitivity and specificity, low detection limits, and signal enhancement achieved through nanomaterial integration.
{"title":"Carboxylated Graphene: An Innovative Approach to Enhanced IgA-SARS-CoV-2 Electrochemical Biosensing.","authors":"Luciana de Souza Freire, Ariamna María Dip Gandarilla, Yonny Romaguera Barcelay, Camila Macena Ruzo, Barbara Batista Salgado, Ana P M Tavares, Francisco Xavier Nobre, Julio Nino de Souza Neto, Spartaco Astolfi-Filho, Ștefan Țălu, Pritesh Lalwani, Niranjan Patra, Walter Ricardo Brito","doi":"10.3390/bios15010034","DOIUrl":"10.3390/bios15010034","url":null,"abstract":"<p><p>Biosensors harness biological materials as receptors linked to transducers, enabling the capture and transformation of primary biorecognition signals into measurable outputs. This study presents a novel carboxylation method for synthesizing carboxylated graphene (CG) under acidic conditions, enhancing biosensing capabilities. The characterization of the CG was performed using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Raman spectroscopy, thermogravimetric analysis (TGA), and X-ray diffraction (XRD). We modified screen-printed carbon electrodes (SPCEs) with CG to immobilize the SARS-CoV-2 N-protein, facilitating targeted detection of IgA antibodies (IgA-SARS-CoV-2). The analytical performance was assessed via electrochemical techniques such as cyclic voltammetry and electrochemical impedance spectroscopy, confirming CG synthesis effectiveness and biosensor functionality. The developed biosensor efficiently detects IgA-SARS-CoV-2 across a dilution range of 1:1000 to 1:200 <i>v</i>/<i>v</i> in a phosphate-buffered saline (PBS) solution, with a limit of detection calculated at 1:1601 <i>v</i>/<i>v</i>. This device shows considerable potential because of its fast response time, miniaturized design facilitated by SPCEs, reduced sample volume requirements, high sensitivity and specificity, low detection limits, and signal enhancement achieved through nanomaterial integration.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11764328/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Prussian Blue (PB) is commonly incorporated into screen-printed enzymatic devices since it enables the determination of the enzymatically produced hydrogen peroxide at low potentials. Inkjet printing is gaining popularity in the development of electrochemical sensors as a substitute for screen printing. This work presents a fully inkjet-printed graphene-Prussian Blue platform, which can be paired with oxidase enzymes to prepare a biosensor of choice. The graphene electrode was inkjet-printed on a flexible polyimide substrate and then thermally and photonically treated with intense pulsed light, followed by inkjet printing of a PB nanoparticle suspension. The optimization of post-printing treatment and electrode deposition conditions was performed to yield a platform with minimal sheet resistance and peak potential differences. A thorough study of PB deposition was conducted: the fully inkjet-printed system was compared against sensors with PB deposited chemically or by drop casting the PB suspension on different kinds of carbon electrodes (glassy carbon, commercial screen-printed, and in-house inkjet-printed electrodes). For hydrogen peroxide detection, the fully inkjet-printed platform exhibits excellent sensitivity, a wider linear range, better linearity, and greater stability towards higher concentrations of peroxide than the other tested electrodes. Finally, lactate oxidase was immobilized in a chitosan matrix, and the prepared biosensor exhibited analytical performance comparable to other lactate sensors found in the literature in a wide, physiologically relevant linear range for measuring lactate concentration in sweat. The development of mediator-modified electrodes with a single fabrication technology, as demonstrated here, paves the way for the scalable production of low-cost, wearable, and flexible biosensors.
{"title":"Fully Inkjet-Printed Flexible Graphene-Prussian Blue Platform for Electrochemical Biosensing.","authors":"Željka Boček, Marko Zubak, Petar Kassal","doi":"10.3390/bios15010028","DOIUrl":"10.3390/bios15010028","url":null,"abstract":"<p><p>Prussian Blue (PB) is commonly incorporated into screen-printed enzymatic devices since it enables the determination of the enzymatically produced hydrogen peroxide at low potentials. Inkjet printing is gaining popularity in the development of electrochemical sensors as a substitute for screen printing. This work presents a fully inkjet-printed graphene-Prussian Blue platform, which can be paired with oxidase enzymes to prepare a biosensor of choice. The graphene electrode was inkjet-printed on a flexible polyimide substrate and then thermally and photonically treated with intense pulsed light, followed by inkjet printing of a PB nanoparticle suspension. The optimization of post-printing treatment and electrode deposition conditions was performed to yield a platform with minimal sheet resistance and peak potential differences. A thorough study of PB deposition was conducted: the fully inkjet-printed system was compared against sensors with PB deposited chemically or by drop casting the PB suspension on different kinds of carbon electrodes (glassy carbon, commercial screen-printed, and in-house inkjet-printed electrodes). For hydrogen peroxide detection, the fully inkjet-printed platform exhibits excellent sensitivity, a wider linear range, better linearity, and greater stability towards higher concentrations of peroxide than the other tested electrodes. Finally, lactate oxidase was immobilized in a chitosan matrix, and the prepared biosensor exhibited analytical performance comparable to other lactate sensors found in the literature in a wide, physiologically relevant linear range for measuring lactate concentration in sweat. The development of mediator-modified electrodes with a single fabrication technology, as demonstrated here, paves the way for the scalable production of low-cost, wearable, and flexible biosensors.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763782/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhaoyang Zhou, Xingyue Chen, Shuang Jiang, Zhuoer Chen, Sixian Wang, Yueyang Ren, Xiaodong Fan, Tao Le
Sulfaquinoxaline (SQX) is widely utilized in aquaculture and animal husbandry due to its broad antimicrobial spectrum and low cost. However, it is difficult to degrade, and there are relevant residues in the aquatic environment, which could be harmful to both the ecological environment and human health. As a new recognition molecule, the aptamer can be recognized with SQX with high affinity and specificity, and the aptamer is no longer adsorbed to AuNPs after binding to SQX, which weakens the catalytic effect of AuNPs. Consequently, an aptasensor for the detection of SQX was successfully developed. This aptasensor exhibits a linear range of 40-640 ng/mL, with a detection limit of 36.95 ng/mL, demonstrating both sensitivity and selectivity. The recoveries of this aptasensor in water samples ranged from 90 to 109.9%, which was quite in line with high-performance liquid chromatography. These findings suggest that the aptasensor is a valuable tool for detecting SQX in aqueous environmental samples.
{"title":"A Label-Free Aptasensor for the Detection of Sulfaquinoxaline Using AuNPs and Aptamer in Water Environment.","authors":"Zhaoyang Zhou, Xingyue Chen, Shuang Jiang, Zhuoer Chen, Sixian Wang, Yueyang Ren, Xiaodong Fan, Tao Le","doi":"10.3390/bios15010030","DOIUrl":"10.3390/bios15010030","url":null,"abstract":"<p><p>Sulfaquinoxaline (SQX) is widely utilized in aquaculture and animal husbandry due to its broad antimicrobial spectrum and low cost. However, it is difficult to degrade, and there are relevant residues in the aquatic environment, which could be harmful to both the ecological environment and human health. As a new recognition molecule, the aptamer can be recognized with SQX with high affinity and specificity, and the aptamer is no longer adsorbed to AuNPs after binding to SQX, which weakens the catalytic effect of AuNPs. Consequently, an aptasensor for the detection of SQX was successfully developed. This aptasensor exhibits a linear range of 40-640 ng/mL, with a detection limit of 36.95 ng/mL, demonstrating both sensitivity and selectivity. The recoveries of this aptasensor in water samples ranged from 90 to 109.9%, which was quite in line with high-performance liquid chromatography. These findings suggest that the aptasensor is a valuable tool for detecting SQX in aqueous environmental samples.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763722/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Li Zheng, Qingdan Ye, Mengmeng Wang, Fan Sun, Qiang Chen, Xiaoping Yu, Yufeng Wang, Pei Liang
Nucleic acid aptamers are single-stranded oligonucleotides that are selected through exponential enrichment (SELEX) technology from synthetic DNA/RNA libraries. These aptamers can specifically recognize and bind to target molecules, serving as specific recognition elements. Surface-enhanced Raman scattering (SERS) spectroscopy is an ultra-sensitive, non-destructive analytical technique that can rapidly acquire the "fingerprint information" of the measured molecules. It has been widely applied in qualitative and trace analysis across various fields, including food safety, environmental monitoring, and biomedical applications. Small molecules, such as toxins, antibiotics, and pesticides, have significant biological effects and are harmful to both human health and the environment. In this paper, we mainly introduced the application and the research progress of SERS detection with aptamers (aptamer-based SERS techniques) in the field of small-molecule detection, particularly in the analysis of pesticide (animal) residues, antibiotics, and toxins. And the progress and prospect of combining the two methods in detection were reviewed.
{"title":"Research Progress in Small-Molecule Detection Using Aptamer-Based SERS Techniques.","authors":"Li Zheng, Qingdan Ye, Mengmeng Wang, Fan Sun, Qiang Chen, Xiaoping Yu, Yufeng Wang, Pei Liang","doi":"10.3390/bios15010029","DOIUrl":"10.3390/bios15010029","url":null,"abstract":"<p><p>Nucleic acid aptamers are single-stranded oligonucleotides that are selected through exponential enrichment (SELEX) technology from synthetic DNA/RNA libraries. These aptamers can specifically recognize and bind to target molecules, serving as specific recognition elements. Surface-enhanced Raman scattering (SERS) spectroscopy is an ultra-sensitive, non-destructive analytical technique that can rapidly acquire the \"fingerprint information\" of the measured molecules. It has been widely applied in qualitative and trace analysis across various fields, including food safety, environmental monitoring, and biomedical applications. Small molecules, such as toxins, antibiotics, and pesticides, have significant biological effects and are harmful to both human health and the environment. In this paper, we mainly introduced the application and the research progress of SERS detection with aptamers (aptamer-based SERS techniques) in the field of small-molecule detection, particularly in the analysis of pesticide (animal) residues, antibiotics, and toxins. And the progress and prospect of combining the two methods in detection were reviewed.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11764255/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
John S Miller, Tanner J Finney, Ethan Ilagan, Skye Frank, Ye Chen-Izu, Keishi Suga, Tonya L Kuhl
Polydiacetylenes (PDAs) are conjugated polymers that are well known for their colorimetric transition from blue to red with the application of energetic stimulus. Sensing platforms based on polymerized diacetylene surfactant vesicles and other structures have been widely demonstrated for various colorimetric biosensing applications. Although less studied and utilized, the transition also results in a change from a non-fluorescent to a highly fluorescent state, making polydiacetylenes useful for both colorimetric and fluorogenic sensing applications. Here, we focus on the characterization and optimization of polydiacetylene vesicles to tune their sensitivity for fluorogenic sensing applications. Particularly, we look at how the structure of the diacetylene (DA) hydrocarbon tail and headgroup affect the self-assembled vesicle size and stability, polymerization kinetics, and the fluorogenic, blue to red phase transition. Longer DA acyl tails generally resulted in smaller and more stable vesicles. The polymerization kinetics and the blue to red transition were a function of both the DA acyl tail length and structure of the headgroup. Decreasing the acyl tail length generally led to vesicles that were more sensitive to energetic stimuli. Headgroup modifications had different effects depending on the structure of the headgroup. Ethanolamine headgroups resulted in vesicles with potentially increased stimuli responsivity. The lower energy stimulus to induce the chromatic transition was attributed to an increase in headgroup hydrogen bonding and polymer backbone strain. Boronic-acid headgroup functionalization led to vesicles that were generally unstable, only weakly polymerized, and unable to fully transform to the red phase due to strong polar, aromatic headgroup interactions. This work presents the design of PDA vesicles in the context of biosensing platforms and includes a discussion of the past, present, and future of PDA biosensing.
{"title":"Fluorogenic Biosensing with Tunable Polydiacetylene Vesicles.","authors":"John S Miller, Tanner J Finney, Ethan Ilagan, Skye Frank, Ye Chen-Izu, Keishi Suga, Tonya L Kuhl","doi":"10.3390/bios15010027","DOIUrl":"10.3390/bios15010027","url":null,"abstract":"<p><p>Polydiacetylenes (PDAs) are conjugated polymers that are well known for their colorimetric transition from blue to red with the application of energetic stimulus. Sensing platforms based on polymerized diacetylene surfactant vesicles and other structures have been widely demonstrated for various colorimetric biosensing applications. Although less studied and utilized, the transition also results in a change from a non-fluorescent to a highly fluorescent state, making polydiacetylenes useful for both colorimetric and fluorogenic sensing applications. Here, we focus on the characterization and optimization of polydiacetylene vesicles to tune their sensitivity for fluorogenic sensing applications. Particularly, we look at how the structure of the diacetylene (DA) hydrocarbon tail and headgroup affect the self-assembled vesicle size and stability, polymerization kinetics, and the fluorogenic, blue to red phase transition. Longer DA acyl tails generally resulted in smaller and more stable vesicles. The polymerization kinetics and the blue to red transition were a function of both the DA acyl tail length and structure of the headgroup. Decreasing the acyl tail length generally led to vesicles that were more sensitive to energetic stimuli. Headgroup modifications had different effects depending on the structure of the headgroup. Ethanolamine headgroups resulted in vesicles with potentially increased stimuli responsivity. The lower energy stimulus to induce the chromatic transition was attributed to an increase in headgroup hydrogen bonding and polymer backbone strain. Boronic-acid headgroup functionalization led to vesicles that were generally unstable, only weakly polymerized, and unable to fully transform to the red phase due to strong polar, aromatic headgroup interactions. This work presents the design of PDA vesicles in the context of biosensing platforms and includes a discussion of the past, present, and future of PDA biosensing.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763271/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143033916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cell lysis is the starting step of many biomedical assays. Electric field-based cell lysis is widely used in many applications, including point-of-care (POC) applications, because it provides an easy one-step solution. Many electric field-based lysis methods utilize micro-electrodes to apply short electric pulses across cells. Unfortunately, these cell lysis devices produce relatively low cell lysis efficiency as electric fields do not reach a significant portion of cells in the sample. Additionally, the utility of syringe pumps for flow cells in and out of the microfluidics channel causes cell loss and low throughput cell lysis. To address these critical issues, we suspended the cells in a sessile drop and concentrated on the electrodes. We used low-frequency AC electric fields (1 Vpp, 0-100 kHz) to drive the cells effectively towards electrodes and lysed using a short pulse of 10 V. A post-lysis analysis was performed using a hemocytometer, UV-vis spectroscopy, and fluorescence imaging. The results show that the pre-electric polarization of cells, prior to applying short electrical pulses, enhances the cell lysis efficiency. Additionally, the application of AC electric fields to concentrate cells on the electrodes reduces the assay time to about 4 min. In this study, we demonstrated that low-frequency AC electric fields can be used to pre-polarize and concentrate cells near micro-electrodes and improve cell lysis efficiency. Due to the simplicity and rapid cell lysis, this method may be suitable for POC assay development.
细胞裂解是许多生物医学检测的第一步。基于电场的细胞裂解技术因其一步到位的简便性而被广泛应用于许多领域,包括护理点(POC)应用。许多基于电场的细胞裂解方法利用微电极在细胞上施加短电脉冲。遗憾的是,这些细胞裂解设备产生的细胞裂解效率相对较低,因为电场无法到达样品中的大部分细胞。此外,使用注射泵使细胞进出微流体通道会造成细胞丢失,从而降低细胞裂解效率。为了解决这些关键问题,我们将细胞悬浮在无柄液滴中,并集中在电极上。我们使用低频交流电场(1 Vpp,0-100 kHz)将细胞有效驱向电极,并使用 10 V 的短脉冲进行裂解。我们使用血细胞计数器、紫外-可见光谱和荧光成像进行了裂解后分析。结果表明,在施加短电脉冲之前对细胞进行预电极化可提高细胞裂解效率。此外,应用交流电场将细胞集中在电极上可将检测时间缩短至约 4 分钟。在这项研究中,我们证明了低频交流电场可用于预极化和集中微电极附近的细胞,并提高细胞裂解效率。由于这种方法简单且细胞裂解速度快,因此可能适用于 POC 检测的开发。
{"title":"The Pre-Polarization and Concentration of Cells near Micro-Electrodes Using AC Electric Fields Enhances the Electrical Cell Lysis in a Sessile Drop.","authors":"Kishor Kaphle, Dharmakeerthi Nawarathna","doi":"10.3390/bios15010022","DOIUrl":"10.3390/bios15010022","url":null,"abstract":"<p><p>Cell lysis is the starting step of many biomedical assays. Electric field-based cell lysis is widely used in many applications, including point-of-care (POC) applications, because it provides an easy one-step solution. Many electric field-based lysis methods utilize micro-electrodes to apply short electric pulses across cells. Unfortunately, these cell lysis devices produce relatively low cell lysis efficiency as electric fields do not reach a significant portion of cells in the sample. Additionally, the utility of syringe pumps for flow cells in and out of the microfluidics channel causes cell loss and low throughput cell lysis. To address these critical issues, we suspended the cells in a sessile drop and concentrated on the electrodes. We used low-frequency AC electric fields (1 Vpp, 0-100 kHz) to drive the cells effectively towards electrodes and lysed using a short pulse of 10 V. A post-lysis analysis was performed using a hemocytometer, UV-vis spectroscopy, and fluorescence imaging. The results show that the pre-electric polarization of cells, prior to applying short electrical pulses, enhances the cell lysis efficiency. Additionally, the application of AC electric fields to concentrate cells on the electrodes reduces the assay time to about 4 min. In this study, we demonstrated that low-frequency AC electric fields can be used to pre-polarize and concentrate cells near micro-electrodes and improve cell lysis efficiency. Due to the simplicity and rapid cell lysis, this method may be suitable for POC assay development.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763957/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lynn Mouawad, Georges Istamboulie, Gaëlle Catanante, Thierry Noguer
A sensitive and reliable electrochemical biosensor for the detection of benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC), the most commonly used disinfectant biocides in the agri-food industry, is described. Acetylcholinesterase from Drosophila melanogaster (DM AChE) and butyrylcholinesterase from horse serum (BChE) were immobilized by entrapment in a photocrosslinkable polymer on the surface of carbon screen-printed electrodes. Preliminary tests conducted in phosphate buffer showed limits of detection (LODs) of 0.26 µM for BAC using the BChE-based sensor and 0.04 µM for DDAC using the DM AChE sensor. These performances comply with the European regulation for dairy products, which sets a maximum allowable concentration of 0.28 µM for biocides. However, when tested directly in milk samples, a dramatic decrease in the sensitivity of both sensors towards BAC and DDAC biocides was reported. To overcome this problem, a simple liquid-liquid extraction was necessary prior to biosensor measurements, ensuring that the biosensors met European regulatory standards and provided an unbiased response.
{"title":"Enhancing Biocide Safety of Milk Using Biosensors Based on Cholinesterase Inhibition.","authors":"Lynn Mouawad, Georges Istamboulie, Gaëlle Catanante, Thierry Noguer","doi":"10.3390/bios15010026","DOIUrl":"10.3390/bios15010026","url":null,"abstract":"<p><p>A sensitive and reliable electrochemical biosensor for the detection of benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC), the most commonly used disinfectant biocides in the agri-food industry, is described. Acetylcholinesterase from <i>Drosophila melanogaster</i> (DM AChE) and butyrylcholinesterase from horse serum (BChE) were immobilized by entrapment in a photocrosslinkable polymer on the surface of carbon screen-printed electrodes. Preliminary tests conducted in phosphate buffer showed limits of detection (LODs) of 0.26 µM for BAC using the BChE-based sensor and 0.04 µM for DDAC using the DM AChE sensor. These performances comply with the European regulation for dairy products, which sets a maximum allowable concentration of 0.28 µM for biocides. However, when tested directly in milk samples, a dramatic decrease in the sensitivity of both sensors towards BAC and DDAC biocides was reported. To overcome this problem, a simple liquid-liquid extraction was necessary prior to biosensor measurements, ensuring that the biosensors met European regulatory standards and provided an unbiased response.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11763983/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143034625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}