The elevated expression of microRNA-21 (miRNA-21) in exosomes derived from early-stage non-small cell lung cancer (NSCLC) holds significant promise for early diagnosis. However, detecting exosomal miRNA-21 remains challenging due to its low abundance. Here, we developed an electrochemiluminescence (ECL) biosensor integrating a 3D DNA walker with rolling circle amplification (RCA) for ultrasensitive miRNA-21 detection. Gold-silver nanoclusters (AuAgNCs, Au:Ag = 4:1) served as ECL emitters, while ferrocene-modified DNA (Fc-DNA) quenched the signal ("signal-off"). Target miRNA-21 activated the 3D DNA walker, generating primers for RCA. The RCA products displaced Fc-DNA, restoring ECL ("signal-on"). The biosensor achieved a wide linear range (1 fM-1 nM) with a 0.14 fM detection limit. Critically, it distinguished NSCLC patients (n = 10) from healthy controls (n = 10) and COPD patients (n = 10) with statistical significance (P < 0.001) in clinical serum samples. Excellent stability (84 % signal retention after 25 days) and reproducibility (RSD = 3.95 %) further support its utility for early lung cancer screening.
{"title":"Bimetallic nanoclusters and dual-amplification tactic: an ultrasensitive electrochemiluminescence biosensor for miRNA-21 detection.","authors":"Tong Shen, Hongwei Yu, Xin Xu, Ze Zhang, Huanzhang Yang, Dong Chang","doi":"10.1016/j.ab.2025.115948","DOIUrl":"10.1016/j.ab.2025.115948","url":null,"abstract":"<p><p>The elevated expression of microRNA-21 (miRNA-21) in exosomes derived from early-stage non-small cell lung cancer (NSCLC) holds significant promise for early diagnosis. However, detecting exosomal miRNA-21 remains challenging due to its low abundance. Here, we developed an electrochemiluminescence (ECL) biosensor integrating a 3D DNA walker with rolling circle amplification (RCA) for ultrasensitive miRNA-21 detection. Gold-silver nanoclusters (AuAgNCs, Au:Ag = 4:1) served as ECL emitters, while ferrocene-modified DNA (Fc-DNA) quenched the signal (\"signal-off\"). Target miRNA-21 activated the 3D DNA walker, generating primers for RCA. The RCA products displaced Fc-DNA, restoring ECL (\"signal-on\"). The biosensor achieved a wide linear range (1 fM-1 nM) with a 0.14 fM detection limit. Critically, it distinguished NSCLC patients (n = 10) from healthy controls (n = 10) and COPD patients (n = 10) with statistical significance (P < 0.001) in clinical serum samples. Excellent stability (84 % signal retention after 25 days) and reproducibility (RSD = 3.95 %) further support its utility for early lung cancer screening.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115948"},"PeriodicalIF":2.5,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144726511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-31DOI: 10.1016/j.ab.2025.116006
Guangxin Yuan , Meng Xia , Ya Li , Honglei Tian , Jianyu Zhu , Yanshuang Wang
Deer placenta and Placenta Hominis are valued traditional Chinese medicines often adulterated with cheaper pig or cow placenta. Current identification methods frequently lack the throughput, speed, or simplicity for effective field monitoring. This study aimed to develop a high-throughput, rapid method to simultaneously identify deer, human, pig, and cow placenta. The approach involved screening a rapid genomic DNA extraction protocol and designing specific primers for the mitochondrial cytochrome b gene of each species, with primers labeled with FAM and Biotin. Following the optimization of PCR amplification conditions, results were visually detected using immunocolloidal gold test strips. The method was rigorously evaluated for its specificity, reproducibility, sensitivity, and stability. The results showed that the extracted DNA was of satisfactory concentration, purity, and integrity. Under optimized conditions (59 °C annealing temperature, 20 cycles), authentic samples for all four placenta types produced two distinct red bands on the test strips, while adulterated and blank controls showed only one control band. Agarose gel electrophoresis confirmed specific amplification for each target with no cross-reactivity. The method demonstrated strong specificity, excellent reproducibility and stability, and a high sensitivity of 0.01 ng μL−1, surpassing conventional electrophoresis. In conclusion, this PCR-based test strip method enables the visual, simultaneous authentication of the four placenta types in a single test, making it highly suitable for on-site rapid monitoring and quality control.
{"title":"A high-throughout PCR test strip method for the rapid identification of four placentas","authors":"Guangxin Yuan , Meng Xia , Ya Li , Honglei Tian , Jianyu Zhu , Yanshuang Wang","doi":"10.1016/j.ab.2025.116006","DOIUrl":"10.1016/j.ab.2025.116006","url":null,"abstract":"<div><div>Deer placenta and Placenta Hominis are valued traditional Chinese medicines often adulterated with cheaper pig or cow placenta. Current identification methods frequently lack the throughput, speed, or simplicity for effective field monitoring. This study aimed to develop a high-throughput, rapid method to simultaneously identify deer, human, pig, and cow placenta. The approach involved screening a rapid genomic DNA extraction protocol and designing specific primers for the mitochondrial <em>cytochrome b</em> gene of each species, with primers labeled with FAM and Biotin. Following the optimization of PCR amplification conditions, results were visually detected using immunocolloidal gold test strips. The method was rigorously evaluated for its specificity, reproducibility, sensitivity, and stability. The results showed that the extracted DNA was of satisfactory concentration, purity, and integrity. Under optimized conditions (59 °C annealing temperature, 20 cycles), authentic samples for all four placenta types produced two distinct red bands on the test strips, while adulterated and blank controls showed only one control band. Agarose gel electrophoresis confirmed specific amplification for each target with no cross-reactivity. The method demonstrated strong specificity, excellent reproducibility and stability, and a high sensitivity of 0.01 ng μL<sup>−1</sup>, surpassing conventional electrophoresis. In conclusion, this PCR-based test strip method enables the visual, simultaneous authentication of the four placenta types in a single test, making it highly suitable for on-site rapid monitoring and quality control.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"709 ","pages":"Article 116006"},"PeriodicalIF":2.5,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145429918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-30DOI: 10.1016/j.ab.2025.116004
Rolf Wibom , David Alsina , Karin Naess , Martin Engvall , Christoph Freyer , Anna Wedell , Anna Wredenberg
We present an optimised luminometric method for measuring muscle mitochondrial ATP production rate (MAPR), adapted to a 96-well microplate format. The enhanced assay enables quantification of ATP production from 12 or more substrate combinations within 15 min, using only 10 μL of isolated mitochondria. The method demonstrates high accuracy and precision, with a validated measurement range of 0.3–70 nmol/min/L. To support clinical interpretation, a reference dataset was established from 92 individuals aged seven months to 79 years. All these individuals were referred for muscle biopsy but were subsequently deemed unlikely to have a mitochondrial disorder following comprehensive clinical evaluation. An overview of the current version of our assays for oxidative phosphorylation (OXPHOS) enzymes is also provided.
As proof of concept, we present three patients carrying pathogenic variants in mitochondrial DNA (ATP6 and MT-TL1) and the nuclear PDHA1 gene. All exhibited decreased MAPR with one or more substrates, along with additional clinical, biochemical, and morphological features consistent with mitochondrial disease.
Furthermore, we illustrate the age-dependent development of MAPR in muscle across the human lifespan, demonstrating a 60–80 % higher maximal capacity for oxidative ATP production in adults compared with young children. In contrast, MAPR supported by fatty acid-derived substrates remains unchanged over the same period.
In conclusion, the improved MAPR assay offers a robust and efficient tool for assessing mitochondrial function in both clinical diagnostics and research. Its high-throughput format and reliable performance make it particularly well-suited for the investigation of suspected mitochondrial disorders.
{"title":"Advancing a sensitive method for measuring mitochondrial ATP production in small muscle biopsy samples","authors":"Rolf Wibom , David Alsina , Karin Naess , Martin Engvall , Christoph Freyer , Anna Wedell , Anna Wredenberg","doi":"10.1016/j.ab.2025.116004","DOIUrl":"10.1016/j.ab.2025.116004","url":null,"abstract":"<div><div>We present an optimised luminometric method for measuring muscle mitochondrial ATP production rate (MAPR), adapted to a 96-well microplate format. The enhanced assay enables quantification of ATP production from 12 or more substrate combinations within 15 min, using only 10 μL of isolated mitochondria. The method demonstrates high accuracy and precision, with a validated measurement range of 0.3–70 nmol/min/L. To support clinical interpretation, a reference dataset was established from 92 individuals aged seven months to 79 years. All these individuals were referred for muscle biopsy but were subsequently deemed unlikely to have a mitochondrial disorder following comprehensive clinical evaluation. An overview of the current version of our assays for oxidative phosphorylation (OXPHOS) enzymes is also provided.</div><div>As proof of concept, we present three patients carrying pathogenic variants in mitochondrial DNA (<em>ATP6</em> and <em>MT-TL1</em>) and the nuclear <em>PDHA1</em> gene. All exhibited decreased MAPR with one or more substrates, along with additional clinical, biochemical, and morphological features consistent with mitochondrial disease.</div><div>Furthermore, we illustrate the age-dependent development of MAPR in muscle across the human lifespan, demonstrating a 60–80 % higher maximal capacity for oxidative ATP production in adults compared with young children. In contrast, MAPR supported by fatty acid-derived substrates remains unchanged over the same period.</div><div>In conclusion, the improved MAPR assay offers a robust and efficient tool for assessing mitochondrial function in both clinical diagnostics and research. Its high-throughput format and reliable performance make it particularly well-suited for the investigation of suspected mitochondrial disorders.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"709 ","pages":"Article 116004"},"PeriodicalIF":2.5,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145420885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-29DOI: 10.1016/j.ab.2025.116002
Katelyn M. Brown , Dianne I. Greenfield , George S. Bullerjahn
A novel, rapid (2.5 h) sandwich hybridization assay (SHA) method for the detection of harmful cyanobacterial genera has been developed based on oligonucleotide hybridization to genus-specific sequences within the 16S ribosomal RNA. Unlike previously-developed 96-well plate SHA formats, this modification does not involve a robotic processor to maneuver reagents across the plate. Rather, it requires manual pipetting each reagent sequentially into the same well and washing the 96-well plate between each addition, thereby increasing sample throughput. Another key difference is the colorimetric reporter molecule. Yet, it enables specific detection of Planktothrix, Raphidiopsis and the Anabaena-Dolichospermum-Aphanizomenon (ADA) clade of the Nostocales. As these are the most common planktonic bloom-forming cyanobacteria, the method can be used to assess potential threats posed by these taxa, specifically cyanotoxins that are associated with particular bloom-forming taxa. The assays detected target groups and standard curves were created from cultured cyanobacteria biomass. Here, the SHA method, calibration, and the field performance of the assay on cyanobacterial bloom biomass is described. Linear standard curves were established for each assay, with a Planktothrix and Raphidiopsis quantification range of 10–500 mm3 L−1 100 μL homogenate−1 and a range of 50–500 mm3 L−1 100 μL homogenate−1 for the ADA clade assay. In environmental samples from three Ohio waterbodies, the assays detected the target genus in a mixed cyanobacterial community. This new SHA protocol will be highly beneficial to both management and research communities because it considerably broadens the capability to rapidly detect multiple cyanobacterial genera that cause persistent toxic blooms.
{"title":"A novel sandwich hybridization assay method and probe sets for cyanobacterial harmful algal bloom detection","authors":"Katelyn M. Brown , Dianne I. Greenfield , George S. Bullerjahn","doi":"10.1016/j.ab.2025.116002","DOIUrl":"10.1016/j.ab.2025.116002","url":null,"abstract":"<div><div>A novel, rapid (2.5 h) sandwich hybridization assay (SHA) method for the detection of harmful cyanobacterial genera has been developed based on oligonucleotide hybridization to genus-specific sequences within the 16S ribosomal RNA. Unlike previously-developed 96-well plate SHA formats, this modification does not involve a robotic processor to maneuver reagents across the plate. Rather, it requires manual pipetting each reagent sequentially into the same well and washing the 96-well plate between each addition, thereby increasing sample throughput. Another key difference is the colorimetric reporter molecule. Yet, it enables specific detection of <em>Planktothrix, Raphidiopsis</em> and the <em>Anabaena-Dolichospermum-Aphanizomenon</em> (ADA) clade of the Nostocales. As these are the most common planktonic bloom-forming cyanobacteria, the method can be used to assess potential threats posed by these taxa, specifically cyanotoxins that are associated with particular bloom-forming taxa. The assays detected target groups and standard curves were created from cultured cyanobacteria biomass. Here, the SHA method, calibration, and the field performance of the assay on cyanobacterial bloom biomass is described. Linear standard curves were established for each assay, with a <em>Planktothrix</em> and <em>Raphidiopsis</em> quantification range of 10–500 mm<sup>3</sup> L<sup>−1</sup> 100 μL homogenate<sup>−1</sup> and a range of 50–500 mm<sup>3</sup> L<sup>−1</sup> 100 μL homogenate<sup>−1</sup> for the ADA clade assay. In environmental samples from three Ohio waterbodies, the assays detected the target genus in a mixed cyanobacterial community. This new SHA protocol will be highly beneficial to both management and research communities because it considerably broadens the capability to rapidly detect multiple cyanobacterial genera that cause persistent toxic blooms.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"709 ","pages":"Article 116002"},"PeriodicalIF":2.5,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145414889","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
We propose a new concept to detect a target molecule on a solid surface directly by spraying biosensing molecules. First, a filter paper as a model of a solid surface was spotted with a target nucleic acid. Then, the biosensing molecule, a DNA-nano-tweezers-structure that exhibits a peroxidase activity recognizing the target, was sprayed with hemin as a cofactor, followed by a second spray containing luminol as a substrate for a light-glowing signal. Finally, an image was recorded using a smartphone. The images revealed that only the places where the corresponding targets were spotted glowed a blue light.
{"title":"Direct detection of a target nucleic acid on a surface by spraying DNA-nano-tweezers-based biosensing molecules","authors":"Hisakage Funabashi, Hiroya Hatano, Hirotaka Nakamura, Reiji Shigematsu, Akio Kuroda","doi":"10.1016/j.ab.2025.116003","DOIUrl":"10.1016/j.ab.2025.116003","url":null,"abstract":"<div><div>We propose a new concept to detect a target molecule on a solid surface directly by spraying biosensing molecules. First, a filter paper as a model of a solid surface was spotted with a target nucleic acid. Then, the biosensing molecule, a DNA-nano-tweezers-structure that exhibits a peroxidase activity recognizing the target, was sprayed with hemin as a cofactor, followed by a second spray containing luminol as a substrate for a light-glowing signal. Finally, an image was recorded using a smartphone. The images revealed that only the places where the corresponding targets were spotted glowed a blue light.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"709 ","pages":"Article 116003"},"PeriodicalIF":2.5,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145407907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-16DOI: 10.1016/j.ab.2025.115994
Lichao Zhang , Xue Wang , Liang Kong
Lysine acetylation (Kace) is an important post-translational modification. Although structure information has been proven to be a key for improving model effectiveness, it is difficult to obtain in bulk due to experiments limitations. In this study, we propose a spatial coordinates representation using a 2-order tensor according to defined property sequence to address the existing limitations and depicting amino acid position in 3-d space. Based on the proposed coordinates, we construct optimal complex networks to extract network-derived features. Compared to existing network construction methods, protein contact networks (PCN), the features achieve superior performance, demonstrating the proposed spatial coordinates could effectively capture biological global information. Meanwhile, we proposed a computational model, named 3DCOOR-Kace, by fusing sequence and structure information based on DenseNet and Squeeze-and-Excitation layer. The 3DCOOR-Kace achieved satisfactory MCC with 0.7358. Compared with MusiteDeep and TransPTM by the independent testing set, the MCC is 0.4261 higher than MusiteDeep and 0.1660 higher than TransPTM, which demonstrates 3DCOOR-Kace are effective for integrating structure and sequence information for improving Kace site identification. Instead of doing biological experiments, the 3-d spatial coordinates representation could give sites positions directly, which could address the experiments limitation and be convenient for computational methods and biological function research.
{"title":"3DCOOR-Kace: A 3-d spatial coordinates representation method for lysine acetylation site identification","authors":"Lichao Zhang , Xue Wang , Liang Kong","doi":"10.1016/j.ab.2025.115994","DOIUrl":"10.1016/j.ab.2025.115994","url":null,"abstract":"<div><div>Lysine acetylation (Kace) is an important post-translational modification. Although structure information has been proven to be a key for improving model effectiveness, it is difficult to obtain in bulk due to experiments limitations. In this study, we propose a spatial coordinates representation using a 2-order tensor according to defined property sequence to address the existing limitations and depicting amino acid position in 3-d space. Based on the proposed coordinates, we construct optimal complex networks to extract network-derived features. Compared to existing network construction methods, protein contact networks (PCN), the features achieve superior performance, demonstrating the proposed spatial coordinates could effectively capture biological global information. Meanwhile, we proposed a computational model, named 3DCOOR-Kace, by fusing sequence and structure information based on DenseNet and Squeeze-and-Excitation layer. The 3DCOOR-Kace achieved satisfactory MCC with 0.7358. Compared with MusiteDeep and TransPTM by the independent testing set, the MCC is 0.4261 higher than MusiteDeep and 0.1660 higher than TransPTM, which demonstrates 3DCOOR-Kace are effective for integrating structure and sequence information for improving Kace site identification. Instead of doing biological experiments, the 3-d spatial coordinates representation could give sites positions directly, which could address the experiments limitation and be convenient for computational methods and biological function research.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"708 ","pages":"Article 115994"},"PeriodicalIF":2.5,"publicationDate":"2025-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145318062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-15DOI: 10.1016/j.ab.2025.115990
Rimpa Mondal , Sk Faruque Ahmed , Nillohit Mukherjee
Carbonaceous materials are placed in the category of advanced and future materials for their very unique physicochemical attributes like tuneable electronic, electrical and optical properties associated with good chemical reactivity, biocompatibility and high surface to volume ratio that make them good candidates for various device applications including biosensors. Recently, such carbonaceous materials and their composites are being widely used as the electrode materials for enzyme-free electrochemical detection of various bioanalytes. Among the bioanalytes, the neurotransmitters are known to play crucial roles in maintaining proper balance in the human nervous systems. However, there are many challenges like selectivity, sensitivity and rapidity; that are normally faced during the detection of such neurotransmitters using the conventional enzymatic way. Electrochemical biosensors made up of advanced carbonaceous materials could be a single-key answer to all such challenges. Serotonin, a catecholamine group neurotransmitter, also known as the “happy hormone”, is known to control human emotions. So, real-time monitoring of its level in human serum carries immense importance, however; it comes with various challenges. This review article covers the developments of advanced carbonaceous, as well as, the 5th generation materials like Mxenes and related composites in the enzyme-free electrochemical sensing of serotonin since the inception of such materials in electrochemical biosensing. A detailed literature review exhibited that, impressive results, like very low limit of detection (LoD) of 8.0 nM could be achieved using multiwalled carbon nanotube (MWCNT) based electrodes, whereas, nitrogen incorporated graphene quantum dot (N-GQD) modified electrodes can help to reach a notably high sensitivity of 24.0 μAμM−1.cm−2. Again, carbon spheres (CS) have been established as a good candidate to bring a balance between LoD (4.0 nM) and sensitivity (5.5 μAμM−1.cm−2) in combination with metal oxide electrode.
{"title":"Electrochemical strategies for decoding serotonin using advanced carbonaceous and 5th generation materials: A comprehensive review","authors":"Rimpa Mondal , Sk Faruque Ahmed , Nillohit Mukherjee","doi":"10.1016/j.ab.2025.115990","DOIUrl":"10.1016/j.ab.2025.115990","url":null,"abstract":"<div><div>Carbonaceous materials are placed in the category of advanced and future materials for their very unique physicochemical attributes like tuneable electronic, electrical and optical properties associated with good chemical reactivity, biocompatibility and high surface to volume ratio that make them good candidates for various device applications including biosensors. Recently, such carbonaceous materials and their composites are being widely used as the electrode materials for enzyme-free electrochemical detection of various bioanalytes. Among the bioanalytes, the neurotransmitters are known to play crucial roles in maintaining proper balance in the human nervous systems. However, there are many challenges like selectivity, sensitivity and rapidity; that are normally faced during the detection of such neurotransmitters using the conventional enzymatic way. Electrochemical biosensors made up of advanced carbonaceous materials could be a single-key answer to all such challenges. Serotonin, a catecholamine group neurotransmitter, also known as the “happy hormone”, is known to control human emotions. So, real-time monitoring of its level in human serum carries immense importance, however; it comes with various challenges. This review article covers the developments of advanced carbonaceous, as well as, the 5th generation materials like Mxenes and related composites in the enzyme-free electrochemical sensing of serotonin since the inception of such materials in electrochemical biosensing. A detailed literature review exhibited that, impressive results, like very low limit of detection (LoD) of 8.0 nM could be achieved using multiwalled carbon nanotube (MWCNT) based electrodes, whereas, nitrogen incorporated graphene quantum dot (N-GQD) modified electrodes can help to reach a notably high sensitivity of 24.0 μAμM<sup>−1</sup>.cm<sup>−2</sup>. Again, carbon spheres (CS) have been established as a good candidate to bring a balance between LoD (4.0 nM) and sensitivity (5.5 μAμM<sup>−1</sup>.cm<sup>−2</sup>) in combination with metal oxide electrode.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"709 ","pages":"Article 115990"},"PeriodicalIF":2.5,"publicationDate":"2025-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145312138","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-14DOI: 10.1016/j.ab.2025.115993
Shweta Mishra , Ashlesha J. Chauhan , Jayaveersinh Mahida
Biosensors have become essential analytical tools that integrate biological recognition elements with physical transducers to detect specific analytes with remarkable sensitivity. This review delves into the applications of biosensors in healthcare, environmental monitoring, and other fields. Notable advancements include nanotechnology-enhanced biosensors, wearable devices, and point-of-care systems, all of which are characterized by miniaturization and multiplexing capabilities. In healthcare, biosensors facilitate disease diagnosis and continuous monitoring, whereas in environmental applications, they aid in pollutant detection. In analytical technology, biosensors merge biological recognition with physical transduction, offering unparalleled sensitivity in biological and environmental systems. This review examines the characteristics and applications of biosensors in various fields. They support disease diagnosis and health surveillance in healthcare and enable real-time pollutant monitoring for environmental protection purposes. Recent advances have incorporated nanotechnology to enhance performance through miniaturization and label-free detection. Wearable biosensors have revolutionized point-of-care diagnostics by providing real-time health data outside clinical settings. The integration of artificial intelligence and microfluidics has led to the development of more adaptive biosensing platforms. Despite challenges such as biological component stability and nonspecific binding, innovations in materials science offer potential solutions. The convergence of expertise positions biosensors as vital instruments for improving health outcomes and ensuring environmental sustainability, thereby transforming diagnostics and ecological preservation.
{"title":"Exploring biosensors: Distinctive features and emerging applications","authors":"Shweta Mishra , Ashlesha J. Chauhan , Jayaveersinh Mahida","doi":"10.1016/j.ab.2025.115993","DOIUrl":"10.1016/j.ab.2025.115993","url":null,"abstract":"<div><div>Biosensors have become essential analytical tools that integrate biological recognition elements with physical transducers to detect specific analytes with remarkable sensitivity. This review delves into the applications of biosensors in healthcare, environmental monitoring, and other fields. Notable advancements include nanotechnology-enhanced biosensors, wearable devices, and point-of-care systems, all of which are characterized by miniaturization and multiplexing capabilities. In healthcare, biosensors facilitate disease diagnosis and continuous monitoring, whereas in environmental applications, they aid in pollutant detection. In analytical technology, biosensors merge biological recognition with physical transduction, offering unparalleled sensitivity in biological and environmental systems. This review examines the characteristics and applications of biosensors in various fields. They support disease diagnosis and health surveillance in healthcare and enable real-time pollutant monitoring for environmental protection purposes. Recent advances have incorporated nanotechnology to enhance performance through miniaturization and label-free detection. Wearable biosensors have revolutionized point-of-care diagnostics by providing real-time health data outside clinical settings. The integration of artificial intelligence and microfluidics has led to the development of more adaptive biosensing platforms. Despite challenges such as biological component stability and nonspecific binding, innovations in materials science offer potential solutions. The convergence of expertise positions biosensors as vital instruments for improving health outcomes and ensuring environmental sustainability, thereby transforming diagnostics and ecological preservation.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"709 ","pages":"Article 115993"},"PeriodicalIF":2.5,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145306919","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-14DOI: 10.1016/j.ab.2025.115992
Gerald A. Dienel
Phenylketonuria (PKU) is an inborn error of metabolism owing to deficits in phenylalanine hydroxylase (PAH) activity. PKU children acquire irreversible brain damage if newborns are not identified and treated with a phenylalanine-restricted diet. In spite of decades of research, the mechanisms underlying PKU brain dysfunction are not adequately understood. Competition of phenylalanine with large neutral amino acids (LNAAs) for carrier-mediated uptake into brain, causing lower brain LNAA levels and reduced neurotransmitter synthesis from tyrosine and tryptophan, is a long-favored mechanism for brain dysfunction. Here, glycine is hypothesized to contribute to phenylalanine-evoked brain disorders. All PKU animal models exhibit elevated brain glycine levels similar to mouse models of nonketotic hyperglycinemia. Glycine is synthesized from l-serine; it is a co-agonist of N-methyl-d-aspartate receptors (NMDARs) and an inhibitory neurotransmitter. l-Serine is synthesized from glucose in astrocytes, exported to neurons, and converted by serine racemase to d-serine, an NMDAR co-agonist. Increased glycine level enhances its inhibition of serine racemase and reduces levels of d-serine. l-Serine-glycine-d-serine interactions can be linked to PAH deficits because elevated brain phenylalanine concentration causes its metabolism by minor pathways to generate phenyllactate. If phenyllactate and l-serine synthesis are coupled via transaminase and redox reactions, the stoichiometry is 1:1. These findings support the following hypothesis: (i) phenylalanine disrupts glycine and d-serine homeostasis during brain maturation, irreversibly altering neuronal development and circuit formation, and (ii) high glycine and low d-serine levels in PKU adults contribute to cognitive and behavioral dysfunction. Suggested new directions for future studies of PKU focus on glycine neurotoxicity.
苯丙酮尿症（PKU）是由于苯丙氨酸羟化酶（PAH）活性不足而导致的先天性代谢错误。如果新生儿未被识别并以苯丙氨酸限制饮食治疗，PKU儿童将获得不可逆的脑损伤。尽管经过数十年的研究，PKU脑功能障碍的机制仍未得到充分的了解。苯丙氨酸与大中性氨基酸（LNAAs）在载体介导的脑摄取中竞争，导致脑LNAA水平降低，酪氨酸和色氨酸合成神经递质减少，这是脑功能障碍的一个长期被认为的机制。在这里，甘氨酸被假设有助于苯丙氨酸诱发的脑部疾病。所有PKU动物模型均表现出脑甘氨酸水平升高，与非酮症型高甘氨酸血症小鼠模型相似。甘氨酸是由l -丝氨酸合成的；它是n -甲基- d -天冬氨酸受体（NMDARs）的协同激动剂和一种抑制性神经递质。l -丝氨酸由星形胶质细胞中的葡萄糖合成，输出到神经元，并通过丝氨酸消旋酶转化为d -丝氨酸，一种NMDAR共激动剂。甘氨酸水平的升高增强了其对丝氨酸消旋酶的抑制作用，降低了d -丝氨酸水平。l -丝氨酸-甘氨酸- d -丝氨酸相互作用可能与多环芳烃缺陷有关，因为脑苯丙氨酸浓度升高导致其通过次要途径代谢产生苯乳酸。如果苯乳酸和l -丝氨酸的合成通过转氨酶和氧化还原反应偶联，则化学计量比为1:1。这些发现支持以下假设：(i)苯丙氨酸破坏脑成熟过程中甘氨酸和d -丝氨酸的稳态，不可逆转地改变神经元发育和电路形成；（ii） PKU成人中高甘氨酸和低d -丝氨酸水平导致认知和行为功能障碍。提出甘氨酸神经毒性是今后研究的新方向。
{"title":"Revisiting phenylketonuria: Do high brain glycine levels caused by chronic hyperphenylalanemia contribute to brain dysfunction by modulating D-serine levels and NMDA receptor activity?","authors":"Gerald A. Dienel","doi":"10.1016/j.ab.2025.115992","DOIUrl":"10.1016/j.ab.2025.115992","url":null,"abstract":"<div><div>Phenylketonuria (PKU) is an inborn error of metabolism owing to deficits in phenylalanine hydroxylase (PAH) activity. PKU children acquire irreversible brain damage if newborns are not identified and treated with a phenylalanine-restricted diet. In spite of decades of research, the mechanisms underlying PKU brain dysfunction are not adequately understood. Competition of phenylalanine with large neutral amino acids (LNAAs) for carrier-mediated uptake into brain, causing lower brain LNAA levels and reduced neurotransmitter synthesis from tyrosine and tryptophan, is a long-favored mechanism for brain dysfunction. Here, glycine is hypothesized to contribute to phenylalanine-evoked brain disorders. All PKU animal models exhibit elevated brain glycine levels similar to mouse models of nonketotic hyperglycinemia. Glycine is synthesized from <span>l</span>-serine; it is a co-agonist of <em>N</em>-methyl-<span>d</span>-aspartate receptors (NMDARs) and an inhibitory neurotransmitter. <span>l</span>-Serine is synthesized from glucose in astrocytes, exported to neurons, and converted by serine racemase to <span>d</span>-serine, an NMDAR co-agonist. Increased glycine level enhances its inhibition of serine racemase and reduces levels of <span>d</span>-serine. <span>l</span>-Serine-glycine-<span>d</span>-serine interactions can be linked to PAH deficits because elevated brain phenylalanine concentration causes its metabolism by minor pathways to generate phenyllactate. If phenyllactate and <span>l</span>-serine synthesis are coupled via transaminase and redox reactions, the stoichiometry is 1:1. These findings support the following hypothesis: (i) phenylalanine disrupts glycine and <span>d</span>-serine homeostasis during brain maturation, irreversibly altering neuronal development and circuit formation, and (ii) high glycine and low <span>d</span>-serine levels in <span>PKU</span> adults contribute to cognitive and behavioral dysfunction. Suggested new directions for future studies of PKU focus on glycine neurotoxicity.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"708 ","pages":"Article 115992"},"PeriodicalIF":2.5,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145306838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-13DOI: 10.1016/j.ab.2025.115991
Leting Yang , Xia Han , Xiao Zhu , Yuhao Wang , Junjiu Xu , Yan Chen , Qiuping Qin , Likun Gong
Accurate measurement of interleukin-8 (IL-8) in non-human primates is crucial for research into inflammation and tumors; however, sensitive enzyme-linked immunosorbent assays (ELISAs) for non-clinical studies remain limited. We aimed to develop and validate a sensitive method for IL-8 quantification in cynomolgus monkey serum. Using two recombinant IL-8-specific antibodies and a surrogate matrix, a sandwich-type ELISA was developed, involving sequential incubations with samples, biotinylated antibody, and horseradish peroxidase-labeled Streptavidin (SA-HRP). The assay was validated against regulatory guidelines, achieving a lower limit of quantification of 4.69 pg/mL as well as good accuracy and precision. The assay was neither affected by serum matrix nor hemolysis, nor did it cross-react with other cytokines. Furthermore, the assay showed dilution linearity, no hook effect, and good recovery and parallelism. Finally, IL-8 in monkey serum exhibited sufficient stability for analysis. After validation, the assay was applied to 19 healthy cynomolgus monkey serum samples, and the results correlated well with those from a commercial assay. In conclusion, the developed ELISA is a sensitive, accurate, and reliable method for IL-8 quantification in cynomolgus monkey serum, suitable for non-clinical studies in cynomolgus monkeys.
{"title":"A sensitive ELISA made with recombinant antibodies for cynomolgus IL-8 in serum","authors":"Leting Yang , Xia Han , Xiao Zhu , Yuhao Wang , Junjiu Xu , Yan Chen , Qiuping Qin , Likun Gong","doi":"10.1016/j.ab.2025.115991","DOIUrl":"10.1016/j.ab.2025.115991","url":null,"abstract":"<div><div>Accurate measurement of interleukin-8 (IL-8) in non-human primates is crucial for research into inflammation and tumors; however, sensitive enzyme-linked immunosorbent assays (ELISAs) for non-clinical studies remain limited. We aimed to develop and validate a sensitive method for IL-8 quantification in cynomolgus monkey serum. Using two recombinant IL-8-specific antibodies and a surrogate matrix, a sandwich-type ELISA was developed, involving sequential incubations with samples, biotinylated antibody, and horseradish peroxidase-labeled Streptavidin (SA-HRP). The assay was validated against regulatory guidelines, achieving a lower limit of quantification of 4.69 pg/mL as well as good accuracy and precision. The assay was neither affected by serum matrix nor hemolysis, nor did it cross-react with other cytokines. Furthermore, the assay showed dilution linearity, no hook effect, and good recovery and parallelism. Finally, IL-8 in monkey serum exhibited sufficient stability for analysis. After validation, the assay was applied to 19 healthy cynomolgus monkey serum samples, and the results correlated well with those from a commercial assay. In conclusion, the developed ELISA is a sensitive, accurate, and reliable method for IL-8 quantification in cynomolgus monkey serum, suitable for non-clinical studies in cynomolgus monkeys.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"708 ","pages":"Article 115991"},"PeriodicalIF":2.5,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145297932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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