Pub Date : 2025-01-08DOI: 10.1016/j.bios.2025.117141
Qianxiu Pan , Hong Guan , Wenjing Xu , Jingjing Zhao , Yan Liu , Lin Cui , Jin Zhou
Chiral isomers show different behaviours and properties in physiological activities. It is of great significance to find productive approach to realize the recognition of enantiomers, which is a key issue in biochemical and pharmaceutical fields. Nowadays, chiral identification can be successfully achieved according to the discrepancies of special signals correlated with different enantiomers of multiple electrode structures. Electrochemical technologies have attracted wide interest in enantioselective analysis because of its unique merits, such as the economic and miniaturized instruments, simplified and environmental-friendly sample preparations. This review summarizes the development trends of electrochemical sensing in the enantiospecific analysis of chiral drugs, expounds the enantiospecific recognition mechanism between chiral selector and target enantiomers based on general electrochemical, electrochemiluminescent and photoelectrochemical sensors, respectively. In addition, this review attempts to predict the future application of electrochemical, electrochemiluminescent and photoelectrochemical-based technologies in the enantioselective recognition and detection.
{"title":"Recent advance for enantiorecognition of chiral drugs sensing: Electrochemical, electrochemiluminescent and photoelectrochemical application","authors":"Qianxiu Pan , Hong Guan , Wenjing Xu , Jingjing Zhao , Yan Liu , Lin Cui , Jin Zhou","doi":"10.1016/j.bios.2025.117141","DOIUrl":"10.1016/j.bios.2025.117141","url":null,"abstract":"<div><div>Chiral isomers show different behaviours and properties in physiological activities. It is of great significance to find productive approach to realize the recognition of enantiomers, which is a key issue in biochemical and pharmaceutical fields. Nowadays, chiral identification can be successfully achieved according to the discrepancies of special signals correlated with different enantiomers of multiple electrode structures. Electrochemical technologies have attracted wide interest in enantioselective analysis because of its unique merits, such as the economic and miniaturized instruments, simplified and environmental-friendly sample preparations. This review summarizes the development trends of electrochemical sensing in the enantiospecific analysis of chiral drugs, expounds the enantiospecific recognition mechanism between chiral selector and target enantiomers based on general electrochemical, electrochemiluminescent and photoelectrochemical sensors, respectively. In addition, this review attempts to predict the future application of electrochemical, electrochemiluminescent and photoelectrochemical-based technologies in the enantioselective recognition and detection.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117141"},"PeriodicalIF":10.7,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143027474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-07DOI: 10.1016/j.bios.2025.117140
Hongzhi Liang , Aori Qileng , Haoran Shen , Ziyi Zhang , Weipeng Liu , Zhen-Lin Xu , Yingju Liu
The development of advanced optical probes for point-of-care testing holds great importance in the field of diagnostic technologies. This study focused on the synthesis of a probe featuring both fluorescent and photothermal responses with single excitation wavelength, which was achieved through the combination of oxidized camellia oleifera shell powder (OC) and Prussian blue nanoparticles (PBNPs). Notably, OC derived from the direct processing of raw material showed fluorescent and phosphorescent emissions simultaneously, and the positions of the two peaks overlapped with the absorbance range of PBNPs, making the fluorescent and phosphorescent emissions of OC effectively quenched by PBNPs. Meanwhile, the photothermal property of PBNPs was activated by the phosphorescent emission of OC through photoinduced energy transfer. As a proof of concept, OC@PBNPs was applied in the dual-channel immunoassay, in which illegal addictive aminopyrine (AP) was chosen as the detection target. Furthermore, a portable device was developed to capture the fluorescent and photothermal signals of OC@PBNPs, rendering the detection method based on OC@PBNPs suitable for point-of-care testing (POCT).
{"title":"Development of fluorescent-photothermal probe based on photoinduced energy transfer: A dual-readout immunosensor for the detection of illegal additive","authors":"Hongzhi Liang , Aori Qileng , Haoran Shen , Ziyi Zhang , Weipeng Liu , Zhen-Lin Xu , Yingju Liu","doi":"10.1016/j.bios.2025.117140","DOIUrl":"10.1016/j.bios.2025.117140","url":null,"abstract":"<div><div>The development of advanced optical probes for point-of-care testing holds great importance in the field of diagnostic technologies. This study focused on the synthesis of a probe featuring both fluorescent and photothermal responses with single excitation wavelength, which was achieved through the combination of oxidized camellia oleifera shell powder (OC) and Prussian blue nanoparticles (PBNPs). Notably, OC derived from the direct processing of raw material showed fluorescent and phosphorescent emissions simultaneously, and the positions of the two peaks overlapped with the absorbance range of PBNPs, making the fluorescent and phosphorescent emissions of OC effectively quenched by PBNPs. Meanwhile, the photothermal property of PBNPs was activated by the phosphorescent emission of OC through photoinduced energy transfer. As a proof of concept, OC@PBNPs was applied in the dual-channel immunoassay, in which illegal addictive aminopyrine (AP) was chosen as the detection target. Furthermore, a portable device was developed to capture the fluorescent and photothermal signals of OC@PBNPs, rendering the detection method based on OC@PBNPs suitable for point-of-care testing (POCT).</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117140"},"PeriodicalIF":10.7,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142982318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-06DOI: 10.1016/j.bios.2025.117138
Xu Yang , Fan Wu , Haowei Huang , Guojun Zheng , Hongyu Zhang , Wenrong Cai , Junyao Li , Zheng-Zhi Yin , Yong Kong
Wearable sensors can easily enable real-time and noninvasive glucose (Glu) monitoring, providing vital information for effectively preventing various complications caused by high glucose level. Here, a wearable sensor based on nanozyme-catalyzed cascade reactions is designed for Glu monitoring in sweat. Au nanoparticles (AuNPs) are anchored to the carbonated zeolitic imidazolate framework-8 (ZIF-8-C), endowing the sensor with Glu oxidase (GOx)-like and peroxidase (POD)-like activity. A flexible screen-printed carbon electrode (SPCE) is decorated with the resultant AuNPs@ZIF-8-C, which is further modified with biocompatible and swellable calcium alginate (CA) gels for the preparation of the wearable Glu sensor. The linear range for Glu detection is 10∼300 μM with a limit of detection (LOD) of 4.99 μM, which covers the physiological Glu concentration range in human sweat (10–200 μM). The developed wearable Glu sensor can fit well with the skin tissues due to the flexibility of the SPCE, and thus it can be successfully applied in real-time and noninvasive monitoring of Glu in human sweat. Additionally, the wearable Glu sensor exhibits high antibacterial activity resulted from the generated hydroxyl radicals (·OH), enabling long-term Glu monitoring in sweat.
{"title":"Au nanoparticles anchored carbonized ZIF-8 for enabling real-time and noninvasive glucose monitoring in sweat","authors":"Xu Yang , Fan Wu , Haowei Huang , Guojun Zheng , Hongyu Zhang , Wenrong Cai , Junyao Li , Zheng-Zhi Yin , Yong Kong","doi":"10.1016/j.bios.2025.117138","DOIUrl":"10.1016/j.bios.2025.117138","url":null,"abstract":"<div><div>Wearable sensors can easily enable real-time and noninvasive glucose (Glu) monitoring, providing vital information for effectively preventing various complications caused by high glucose level. Here, a wearable sensor based on nanozyme-catalyzed cascade reactions is designed for Glu monitoring in sweat. Au nanoparticles (AuNPs) are anchored to the carbonated zeolitic imidazolate framework-8 (ZIF-8-C), endowing the sensor with Glu oxidase (GOx)-like and peroxidase (POD)-like activity. A flexible screen-printed carbon electrode (SPCE) is decorated with the resultant AuNPs@ZIF-8-C, which is further modified with biocompatible and swellable calcium alginate (CA) gels for the preparation of the wearable Glu sensor. The linear range for Glu detection is 10∼300 μM with a limit of detection (LOD) of 4.99 μM, which covers the physiological Glu concentration range in human sweat (10–200 μM). The developed wearable Glu sensor can fit well with the skin tissues due to the flexibility of the SPCE, and thus it can be successfully applied in real-time and noninvasive monitoring of Glu in human sweat. Additionally, the wearable Glu sensor exhibits high antibacterial activity resulted from the generated hydroxyl radicals (·OH), enabling long-term Glu monitoring in sweat.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"272 ","pages":"Article 117138"},"PeriodicalIF":10.7,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142942019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-04DOI: 10.1016/j.bios.2025.117134
Yuxin Zhang , Jiahao Zheng , Bayinqiaoge , Tim Cole , Chengchen Zhang , Yi Wang , Shi-Yang Tang
Closed-channel microfluidic systems offer versatile on-chip capabilities for bioanalysis but often face complex fabrication and operational challenges. In contrast, free-boundary off-chip microfluidic platforms are relatively simple to fabricate and operate but lack the ability to perform complex tasks such as on-demand single-target sorting and encapsulation. To address these challenges, we develop an off-chip platform powered by a fluorescent-activated mechanical droplet sorting and production (FAM-DSP) system. The system integrates target detection, sorting, encapsulation, and on-demand droplet generation into a single compact platform, eliminating the need for microfabrication and minimizing the use of specialized fluidic control equipment. It achieves precise single-target encapsulation with a high efficiency of over 70%. Such a capability is applied for improving the performance of droplet digital enzyme-linked immunosorbent assay (ddELISA) by reducing the number of empty droplets and increasing the throughput, enabling precise quantification of target biomarkers with a low limit of detection. This versatile off-chip platform holds promise not only for biomarker detection but also for single-cell analysis and various applications in clinical diagnostics and biomedical research.
{"title":"An off-chip platform for on-demand, single-target encapsulation for ultrasensitive biomarker detection","authors":"Yuxin Zhang , Jiahao Zheng , Bayinqiaoge , Tim Cole , Chengchen Zhang , Yi Wang , Shi-Yang Tang","doi":"10.1016/j.bios.2025.117134","DOIUrl":"10.1016/j.bios.2025.117134","url":null,"abstract":"<div><div>Closed-channel microfluidic systems offer versatile on-chip capabilities for bioanalysis but often face complex fabrication and operational challenges. In contrast, free-boundary off-chip microfluidic platforms are relatively simple to fabricate and operate but lack the ability to perform complex tasks such as on-demand single-target sorting and encapsulation. To address these challenges, we develop an off-chip platform powered by a fluorescent-activated mechanical droplet sorting and production (FAM-DSP) system. The system integrates target detection, sorting, encapsulation, and on-demand droplet generation into a single compact platform, eliminating the need for microfabrication and minimizing the use of specialized fluidic control equipment. It achieves precise single-target encapsulation with a high efficiency of over 70%. Such a capability is applied for improving the performance of droplet digital enzyme-linked immunosorbent assay (ddELISA) by reducing the number of empty droplets and increasing the throughput, enabling precise quantification of target biomarkers with a low limit of detection. This versatile off-chip platform holds promise not only for biomarker detection but also for single-cell analysis and various applications in clinical diagnostics and biomedical research.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"272 ","pages":"Article 117134"},"PeriodicalIF":10.7,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142942002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-04DOI: 10.1016/j.bios.2025.117135
Yao Yue , Min Liu , Mingyi Ma , Zhihao Xu , Haoda Zhang , Qingxiang Wang , Ruijiang Liu
To enhance the biomarker diagnostics sensitivity and selectivity of human papillomavirus type 16 oncoprotein E7 (HPV16 E7) in serum, a label/enzyme-free electrochemical detection platform was developed. This platform featured a type of "Super-turn-off" nanobiosensor monitored through differential pulse voltammetry (DPV). It integrated the magnetic self-assembly property of the α-Fe2O3/Fe3O4@Au/Sub/BSA signal transport nano-medium with the high specificity of CRISPR/Cas14a and the amplification capability of the bipedal walker (DNA walker composed of two ssDNA strands), resulting in the enhanced specificity and anti-interference performance while remaining stable at 4 °C for over 30 days. The results demonstrated that the combination of walker and CRISPR yielded superior sensitivity and analytical capability compared with using either technology alone, achieving a detection limit of 67.17 fg mL−1, a quantification limit of 0.22 pg mL−1, and serum sample recovery rates of 98.46%–115.78%. Additionally, this platform was applied to detect serum and tissue samples from mouse models at various stages of cervical cancer, significantly improving the accuracy and effectiveness of early diagnosis and prognostic evaluation. This novel approach held promise as an efficient tool for point-of-care clinical detection of HPV16 E7, potentially reducing cervical cancer mortality.
{"title":"CRISPR/Cas14a integrated with DNA walker based on magnetic self-assembly for human papillomavirus type 16 oncoprotein E7 ultrasensitive detection","authors":"Yao Yue , Min Liu , Mingyi Ma , Zhihao Xu , Haoda Zhang , Qingxiang Wang , Ruijiang Liu","doi":"10.1016/j.bios.2025.117135","DOIUrl":"10.1016/j.bios.2025.117135","url":null,"abstract":"<div><div>To enhance the biomarker diagnostics sensitivity and selectivity of human papillomavirus type 16 oncoprotein E7 (HPV16 E7) in serum, a label/enzyme-free electrochemical detection platform was developed. This platform featured a type of \"Super-turn-off\" nanobiosensor monitored through differential pulse voltammetry (DPV). It integrated the magnetic self-assembly property of the α-Fe<sub>2</sub>O<sub>3</sub>/Fe<sub>3</sub>O<sub>4</sub>@Au/Sub/BSA signal transport nano-medium with the high specificity of CRISPR/Cas14a and the amplification capability of the bipedal walker (DNA walker composed of two ssDNA strands), resulting in the enhanced specificity and anti-interference performance while remaining stable at 4 °C for over 30 days. The results demonstrated that the combination of walker and CRISPR yielded superior sensitivity and analytical capability compared with using either technology alone, achieving a detection limit of 67.17 fg mL<sup>−1</sup>, a quantification limit of 0.22 pg mL<sup>−1</sup>, and serum sample recovery rates of 98.46%–115.78%. Additionally, this platform was applied to detect serum and tissue samples from mouse models at various stages of cervical cancer, significantly improving the accuracy and effectiveness of early diagnosis and prognostic evaluation. This novel approach held promise as an efficient tool for point-of-care clinical detection of HPV16 E7, potentially reducing cervical cancer mortality.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"272 ","pages":"Article 117135"},"PeriodicalIF":10.7,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142997217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-04DOI: 10.1016/j.bios.2025.117132
Shiliang Bi, Chunwang Chai, Hanxiao He, Yang Zhao
Abnormal levels of thrombin may be associated with various diseases, such as thrombosis and hemorrhagic diseases, making precise detection of thrombin particularly important. Dual signal detection is a method that enhances detection sensitivity and specificity by simultaneously utilizing two different signals. Its primary advantages include improving detection accuracy and reducing false positive rates, making it particularly suitable for clinical analysis and diagnostics. In this work, we developed a dual signal detection method for thrombin based on DNA self-assembly. This design incorporates an X-DNA structure. The two bottom arms of the X-shaped DNA (X-DNA) are designed to bind to CuInS2 nanoparticles via dehydration reactions between amine and carboxyl groups. The two top arms of the X-DNA are designed to hybridize with complementary DNA/glucose oxidase (GOx) and DNA/ferrocene (Fc), respectively. Thrombin triggers the hybridization of DNA/GOx and X-DNA, simultaneously causing the dissociation of DNA/Fc from X-DNA. In the Photoelectrochemical mode, GOx can react with O2 and glucose in the detection solution, resulting in a corresponding decrease in the amount of O2 acting as an electron acceptor and a decrease in the photoelectric signal. In the Differential Pulse Voltammetry mode, due to the decrease in Fc content, the DPV signal also shows a weakening trend. The detection method exhibits a good linear relationship within the range of 10 fM −10 nM, with a detection limit of 6.89 fM and 5.86 fM. The enhanced analytical sensitivity and specificity of dual signal detection technology offer broad prospects for improving disease diagnosis.
凝血酶水平异常可能与多种疾病有关，如血栓形成和出血性疾病，因此凝血酶的精确检测尤为重要。双信号检测是一种通过同时利用两种不同的信号来提高检测灵敏度和特异性的方法。它的主要优点包括提高检测准确性和减少假阳性率，使其特别适用于临床分析和诊断。在这项工作中，我们开发了一种基于DNA自组装的凝血酶双信号检测方法。这个设计包含了一个X-DNA结构。x形DNA （X-DNA）的两个底部臂被设计为通过胺和羧基之间的脱水反应与CuInS2纳米颗粒结合。X-DNA的两个上臂分别用于与互补DNA/葡萄糖氧化酶（GOx）和DNA/二茂铁（Fc）杂交。凝血酶触发DNA/GOx和X-DNA的杂交，同时引起DNA/Fc与X-DNA的分离。在光电化学模式下，GOx可以与检测溶液中的O2和葡萄糖发生反应，导致作为电子受体的O2数量相应减少，光电信号减弱。在差分脉冲伏安模式下，由于Fc含量的降低，DPV信号也呈现出减弱的趋势。检测方法在10 fM -10 nM范围内呈良好的线性关系，检测限分别为6.89 fM和5.86 fM。双信号检测技术提高了分析灵敏度和特异性，为改善疾病诊断提供了广阔的前景。
{"title":"Ultrasensitive dual-mode biosensor for photoelectrochemical and differential pulse voltammetry detection of thrombin based on DNA self-assembly","authors":"Shiliang Bi, Chunwang Chai, Hanxiao He, Yang Zhao","doi":"10.1016/j.bios.2025.117132","DOIUrl":"10.1016/j.bios.2025.117132","url":null,"abstract":"<div><div>Abnormal levels of thrombin may be associated with various diseases, such as thrombosis and hemorrhagic diseases, making precise detection of thrombin particularly important. Dual signal detection is a method that enhances detection sensitivity and specificity by simultaneously utilizing two different signals. Its primary advantages include improving detection accuracy and reducing false positive rates, making it particularly suitable for clinical analysis and diagnostics. In this work, we developed a dual signal detection method for thrombin based on DNA self-assembly. This design incorporates an X-DNA structure. The two bottom arms of the X-shaped DNA (X-DNA) are designed to bind to CuInS<sub>2</sub> nanoparticles via dehydration reactions between amine and carboxyl groups. The two top arms of the X-DNA are designed to hybridize with complementary DNA/glucose oxidase (GOx) and DNA/ferrocene (Fc), respectively. Thrombin triggers the hybridization of DNA/GOx and X-DNA, simultaneously causing the dissociation of DNA/Fc from X-DNA. In the Photoelectrochemical mode, GOx can react with O<sub>2</sub> and glucose in the detection solution, resulting in a corresponding decrease in the amount of O<sub>2</sub> acting as an electron acceptor and a decrease in the photoelectric signal. In the Differential Pulse Voltammetry mode, due to the decrease in Fc content, the DPV signal also shows a weakening trend. The detection method exhibits a good linear relationship within the range of 10 fM −10 nM, with a detection limit of 6.89 fM and 5.86 fM. The enhanced analytical sensitivity and specificity of dual signal detection technology offer broad prospects for improving disease diagnosis.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"272 ","pages":"Article 117132"},"PeriodicalIF":10.7,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142997342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-04DOI: 10.1016/j.bios.2025.117137
Bei Yuan , Zhiqiang Li , Peiwu Li , Qi Zhang , Qingqing Yang , Xiaoqian Tang
There is a phenomenon of combined contamination of fungal toxins, of which aflatoxin B1 (AFB1) is the most toxic, and deoxynivalenol (DON) contamination is common. The use of antigens for double or multiple testing of mycotoxins is easy to cause environmental pollution, and surrogate antigens have become necessary. The small molecule and susceptibility to genetic modification of nanobodies can be used to develop alternative antigens for mycotoxins. In this study, using the nanobody gene sequences of the heavy chain recognition regions of anti-aflatoxin and deoxynivalenol monoclonal antibodies, recombinant plasmids were successfully constructed by one-step cloning, and low-temperature-induced bispecific nanobodies against AFB1-DON were obtained, which can be used as alternative antigens to reduce the pollution of the environment from mycotoxin detection. Enzyme-linked immunosorbent assay validated the bispecific nanobody, and the semi-inhibitory concentration (IC50) of the bispecific nanobody were 0.47 μg/L and 149 μg/L for AFB1 and DON, respectively. Finally, a time-resolved fluorescent dual-detection test strip was constructed by this bispecific nanobody as a surrogate antigen for AFB1 and DON, which was capable of detecting AFB1 and DON at the same time, and the limits of detection (LOD) for the two toxins were 0.0254 μg/L and 21.4 μg/L, respectively. This method has satisfactory sensitivity and does not require antigen, which reduces the toxicity of using antigen.
{"title":"Genetically engineered integrated aflatoxin B1 and deoxynivalenol bispecific nanobody as surrogate antigens for constructed time-resolved immunoassay dual detection methods","authors":"Bei Yuan , Zhiqiang Li , Peiwu Li , Qi Zhang , Qingqing Yang , Xiaoqian Tang","doi":"10.1016/j.bios.2025.117137","DOIUrl":"10.1016/j.bios.2025.117137","url":null,"abstract":"<div><div>There is a phenomenon of combined contamination of fungal toxins, of which aflatoxin B<sub>1</sub> (AFB<sub>1</sub>) is the most toxic, and deoxynivalenol (DON) contamination is common. The use of antigens for double or multiple testing of mycotoxins is easy to cause environmental pollution, and surrogate antigens have become necessary. The small molecule and susceptibility to genetic modification of nanobodies can be used to develop alternative antigens for mycotoxins. In this study, using the nanobody gene sequences of the heavy chain recognition regions of anti-aflatoxin and deoxynivalenol monoclonal antibodies, recombinant plasmids were successfully constructed by one-step cloning, and low-temperature-induced bispecific nanobodies against AFB<sub>1</sub>-DON were obtained, which can be used as alternative antigens to reduce the pollution of the environment from mycotoxin detection. Enzyme-linked immunosorbent assay validated the bispecific nanobody, and the semi-inhibitory concentration (IC<sub>50</sub>) of the bispecific nanobody were 0.47 μg/L and 149 μg/L for AFB<sub>1</sub> and DON, respectively. Finally, a time-resolved fluorescent dual-detection test strip was constructed by this bispecific nanobody as a surrogate antigen for AFB<sub>1</sub> and DON, which was capable of detecting AFB<sub>1</sub> and DON at the same time, and the limits of detection (LOD) for the two toxins were 0.0254 μg/L and 21.4 μg/L, respectively. This method has satisfactory sensitivity and does not require antigen, which reduces the toxicity of using antigen.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"273 ","pages":"Article 117137"},"PeriodicalIF":10.7,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142982321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-04DOI: 10.1016/j.bios.2025.117136
Meltem Agar , Maisem Laabei , Hannah S. Leese , Pedro Estrela
Bacteria pose a significant threat to human health as they can cause diseases and outbreaks; therefore rapid, easy, and specific detection of bacteria in a short time is crucial. Various methods such as polymerase chain reaction and enzyme-linked immunosorbent assay have been developed for bacteria detection. However, most of these methods require sample preparation, trained personnel, and 2–4 days for identification. In this study, an electrochemical sensor has been developed in which a molecularly imprinted polymer (MIP) and aptamer were used together as a bioreceptor for the multiplexed detection of Staphylococcus aureus and Escherichia coli. Non-Faradaic electrochemical impedance spectroscopy (EIS) was employed to assess bacterial detection. Sensor performance was assessed in buffer solution, deionized water and spiked tap water. Aptamer-molecularly imprinted polymer (Apta-MIP) based electrochemical sensors demonstrate high sensitivity and selectivity for the detection of S. aureus and E. coli, with limits of detection of 4 CFU/mL and 2 CFU/mL, respectively. Additionally, these sensors exhibited a broad dynamic range from 1 CFU/mL to 108 CFU/mL. The Apta-MIPs performance surpasses those obtained for Aptasensors alone and MIPs alone, demonstrating the high efficiency of the double recognition effect that originates from the affinity between aptamer and bacteria and target-specific cavities on the polymer. This is the first study in which aptamers and imprinted polymers were used as a hybrid bioreceptors for multiplexed detection of bacteria. The Apta-MIP sensors produced in this study can be used as a point-of-care diagnostic tool for bacteria-related diseases and test of water quality.
{"title":"Aptamer-molecularly imprinted polymer sensors for the detection of bacteria in water","authors":"Meltem Agar , Maisem Laabei , Hannah S. Leese , Pedro Estrela","doi":"10.1016/j.bios.2025.117136","DOIUrl":"10.1016/j.bios.2025.117136","url":null,"abstract":"<div><div>Bacteria pose a significant threat to human health as they can cause diseases and outbreaks; therefore rapid, easy, and specific detection of bacteria in a short time is crucial. Various methods such as polymerase chain reaction and enzyme-linked immunosorbent assay have been developed for bacteria detection. However, most of these methods require sample preparation, trained personnel, and 2–4 days for identification. In this study, an electrochemical sensor has been developed in which a molecularly imprinted polymer (MIP) and aptamer were used together as a bioreceptor for the multiplexed detection of <em>Staphylococcus aureus</em> and <em>Escherichia coli.</em> Non-Faradaic electrochemical impedance spectroscopy (EIS) was employed to assess bacterial detection. Sensor performance was assessed in buffer solution, deionized water and spiked tap water. Aptamer-molecularly imprinted polymer (Apta-MIP) based electrochemical sensors demonstrate high sensitivity and selectivity for the detection of <em>S. aureus</em> and <em>E</em>. <em>coli</em>, with limits of detection of 4 CFU/mL and 2 CFU/mL, respectively. Additionally, these sensors exhibited a broad dynamic range from 1 CFU/mL to 10<sup>8</sup> CFU/mL. The Apta-MIPs performance surpasses those obtained for Aptasensors alone and MIPs alone, demonstrating the high efficiency of the double recognition effect that originates from the affinity between aptamer and bacteria and target-specific cavities on the polymer. This is the first study in which aptamers and imprinted polymers were used as a hybrid bioreceptors for multiplexed detection of bacteria. The Apta-MIP sensors produced in this study can be used as a point-of-care diagnostic tool for bacteria-related diseases and test of water quality.</div></div>","PeriodicalId":259,"journal":{"name":"Biosensors and Bioelectronics","volume":"272 ","pages":"Article 117136"},"PeriodicalIF":10.7,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142963373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-04DOI: 10.1016/j.bios.2025.117130
Xiang-Fei Li, Fu-Gen Wu
The cellular microenvironment exerts a pivotal regulatory influence on cell survival, function, and behavior. Dynamic analysis and detection of the cellular microenvironment can promptly elucidate changes in cellular microenvironmental information, uncover the pathogenesis of diseases associated with aberrant microenvironments, and aid in predicting disease risk and monitoring disease progression. Aggregation-induced emission (AIE) fluorescent molecules possess unique AIE characteristics and offer significant advantages in imaging and sensing cellular microenvironments. In this review, we present a profile of the remarkable progress achieved in utilizing AIE fluorescent molecules for detecting cellular microenvironments in recent years. We particularly focus on AIE fluorescent probes applied in imaging key parameters of the cellular microenvironment, including pH, viscosity, polarity, and temperature, as well as in analyzing critical biological components of the microenvironment, such as gas signal molecules, metal ions, redox state, and proteins. We underscore the design principles, detection mechanisms, sensing performance, and biological applications of these fluorescent probes. Furthermore, we address the current challenges confronting this field and provide prospects for the future development of AIE probes used for microenvironment detection. We trust that this review will inspire researchers to develop more precise and sensitive AIE fluorescent probes for the detection of cellular microenvironments.
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Pub Date : 2025-01-03DOI: 10.1016/j.bios.2025.117131
Mohamed Ibrahim Halawa , Fathalla Belal , Alaa A. Salem , Lei Su , Xueji Zhang
Highly ordered ultrathin nanosheets (NSs) of Au(I)-Cys were fabricated through aggregation-induced supramolecular self-assembly triggered by an extended agitation in an alkaline environment. The synthesized Au(I)-Cys NSs exhibited intense luminescence and exceptional chirality. Remarkably, additions of biothiols to Au(I)-Cys NSs have significantly enhanced their luminescence emission, and circular dichroism properties coupled with morphological modulations into nanoflowers, nanodendrites, or closely packed aggregates. These new findings of Aggregation-Induced Luminescence Enhancement (AIEE) and Aggregation-Induced Circular Dichroism Enhancement (AICE) were attributed to multiple interactions involved such as Au-S bonding, stacked H-bonding, and strong aurophilic Au(I)···Au(I), ligand-metal-charge-transfer (LMCT) and ligand-metal-metal-charge-transfer (LMMCT). The AIEE phenomenon of the fabricated Au(I)-Cys NSs was utilized for developing a highly sensitive luminescent platform for determining homocysteine (Hcy), cysteine (Cys), and glutathione (GSH) biothiols in human serum. The developed platform is simple, fast, sensitive, and highly selective for the determination of biothiols through the concentration ranges of (0.25–100.0 μM), (0.625–40.0 μM), and (5.00–600.0 μM), with a lower detection limit (S/N = 3:1) of 0.15, 0.10 and 1.20 μM for Hcy, Cys, and GSH; respectively. Interestingly, irradiation of Au(I)-Cys NSs with a high-energy electron beam during TEM analysis led to an in-situ transformation of the Au(I)-Cys NSs into gold nanoclusters (AuNCs). This phenomenon provided an innovative bottom-up strategy for the synthesis of AuNCs that could be employed in various biological and therapeutic applications. Optimization of the applied voltage and electron beam's exposure time has been found effective in synthesizing precisely designed and size-controlled AuNCs.
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