Pub Date : 2024-11-15DOI: 10.1016/j.bioelechem.2024.108848
Aleksandra Stefanowska , Michał Czapczyński , Piotr Koprowski , Adam Szewczyk , Paweł Krysiński
The influence of incorporation of mitochondrial inner membrane potassium channel, and channel-forming peptide – Gramicidin on the ion transport and electromechanical properties of model lipid membranes tethered to gold electrode was electrochemically investigated by chronoamperometric and impedance spectroscopy techniques. In the case of the potassium channel the ion transport properties were modulated with channel-specific inhibitor – ATP-Mg2+ complex, whereas in the case of gramicidin peptide – by replacing potassium with sodium ions. The observed two exponential current–time responses of the systems studied were interpreted in terms of ion penetration and electrostriction of tethered lipid bilayer membrane, and conclusions supported with the experiments on alkanethiol self-assembled monolayers of different alkanethiol chain lengths deposited on gold.
{"title":"Time-resolved electromechanical and conductive behavior of nanostructured bilayers tethered to the surface of the electrode with incorporated channel proteins and peptides","authors":"Aleksandra Stefanowska , Michał Czapczyński , Piotr Koprowski , Adam Szewczyk , Paweł Krysiński","doi":"10.1016/j.bioelechem.2024.108848","DOIUrl":"10.1016/j.bioelechem.2024.108848","url":null,"abstract":"<div><div>The influence of incorporation of mitochondrial inner membrane potassium channel, and channel-forming peptide – Gramicidin on the ion transport and electromechanical properties of model lipid membranes tethered to gold electrode was electrochemically investigated by chronoamperometric and impedance spectroscopy techniques. In the case of the potassium channel the ion transport properties were modulated with channel-specific inhibitor – ATP-Mg<sup>2+</sup> complex, whereas in the case of gramicidin peptide – by replacing potassium with sodium ions. The observed two exponential current–time responses of the systems studied were interpreted in terms of ion penetration and electrostriction of tethered lipid bilayer membrane, and conclusions supported with the experiments on alkanethiol self-assembled monolayers of different alkanethiol chain lengths deposited on gold.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"162 ","pages":"Article 108848"},"PeriodicalIF":4.8,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142674657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-05DOI: 10.1016/j.bioelechem.2024.108845
Haotian Xie , Zhaojiang Yin , Guobin Wei , Binghui Li , Hanfeng Cui , Hao Fan , Jing Zhang
This study developed a dual-mode “on-off-on” sensor based on a bipyridine ruthenium metal–organic framework (Ru-MOF) and dual enzyme cleavage technology for the sensitive detection of the K-ras gene. The sensor combines electrogenerated chemiluminescence (ECL) and fluorescence (FL) detection modes, achieving high sensitivity and specificity in detecting the K-ras gene through catalytic hairpin assembly (CHA) and dual enzyme cleavage reactions. Experimental results showed that the detection limits for the K-ras gene were 0.044 fM (ECL) and 0.16 fM (FL), demonstrating excellent selectivity and stability during detection. Through testing actual samples, the sensor has shown potential for application in complex biological environments. This method offers an efficient and reliable new tool for cancer diagnosis and treatment.
{"title":"Sensitive detection of K-ras gene by a dual-mode “on-off-on” sensor based on bipyridine ruthenium-MOF and bis-enzymatic cleavage technology","authors":"Haotian Xie , Zhaojiang Yin , Guobin Wei , Binghui Li , Hanfeng Cui , Hao Fan , Jing Zhang","doi":"10.1016/j.bioelechem.2024.108845","DOIUrl":"10.1016/j.bioelechem.2024.108845","url":null,"abstract":"<div><div>This study developed a dual-mode “on-off-on” sensor based on a bipyridine ruthenium metal–organic framework (Ru-MOF) and dual enzyme cleavage technology for the sensitive detection of the K-ras gene. The sensor combines electrogenerated chemiluminescence (ECL) and fluorescence (FL) detection modes, achieving high sensitivity and specificity in detecting the K-ras gene through catalytic hairpin assembly (CHA) and dual enzyme cleavage reactions. Experimental results showed that the detection limits for the K-ras gene were 0.044 fM (ECL) and 0.16 fM (FL), demonstrating excellent selectivity and stability during detection. Through testing actual samples, the sensor has shown potential for application in complex biological environments. This method offers an efficient and reliable new tool for cancer diagnosis and treatment.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108845"},"PeriodicalIF":4.8,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-02DOI: 10.1016/j.bioelechem.2024.108844
Yaping Zhang , Haiyan Wei , Liang Guo , Wei Gao , Di Cheng , Yanju Liu
The KRAS G12C mutations, as crucial biomarkers, are closely associated with non-small cell lung cancer. Here, a novel label-free electrochemical biosensor with synergistic signal amplification of photocell energy transfer-reversible addition fragmentation chain transfer (PET-RAFT) and ring-opening polymerization (ROP) was developed for the first time for sensitive detection of KRAS G12C mutations. Specifically, hairpin DNA (hDNA), which act as biomolecular probe, was self-assembled on Au electrode surface by Au-S bond. 4-cyano-4-[(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid (CDTPA), the chain transfer agent of PET-RAFT reaction, was then attached to hDNA via amide bond. After that, the target DNA (tDNA) was captured on the electrode surface by complementary base pairing with hDNA. Subsequently, large numbers of electro-active monomers N-acryloxysuccinimide (NAS) were successfully grafted to the electrode surface via PET-RAFT reaction, which provided plenty of junction sites for doxorubicin-polycaprolactone (Dox-PCL) synthesized by ROP. Finally, the Dox-PCL was connected to the electrode surface by ester bond, significantly amplifying the electrochemical signal. Under optimized conditions, the biosensor has a wide linear detection range of 0.1 pM to 1 μM, with a detection limit of 86.9 fM. Attribute to its high sensitivity, specificity, reproducibility and stability, this biosensor possesses considerable potential in early diagnosis of disease and biomedical research.
{"title":"A novel label-free impedance biosensor for KRAS G12C mutations detection based on PET-RAFT and ROP synergistic signal amplification","authors":"Yaping Zhang , Haiyan Wei , Liang Guo , Wei Gao , Di Cheng , Yanju Liu","doi":"10.1016/j.bioelechem.2024.108844","DOIUrl":"10.1016/j.bioelechem.2024.108844","url":null,"abstract":"<div><div>The KRAS G12C mutations, as crucial biomarkers, are closely associated with non-small cell lung cancer. Here, a novel label-free electrochemical biosensor with synergistic signal amplification of photocell energy transfer-reversible addition fragmentation chain transfer (PET-RAFT) and ring-opening polymerization (ROP) was developed for the first time for sensitive detection of KRAS G12C mutations. Specifically, hairpin DNA (hDNA), which act as biomolecular probe, was self-assembled on Au electrode surface by Au-S bond. 4-cyano-4-[(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid (CDTPA), the chain transfer agent of PET-RAFT reaction, was then attached to hDNA via amide bond. After that, the target DNA (tDNA) was captured on the electrode surface by complementary base pairing with hDNA. Subsequently, large numbers of electro-active monomers N-acryloxysuccinimide (NAS) were successfully grafted to the electrode surface via PET-RAFT reaction, which provided plenty of junction sites for doxorubicin-polycaprolactone (Dox-PCL) synthesized by ROP. Finally, the Dox-PCL was connected to the electrode surface by ester bond, significantly amplifying the electrochemical signal. Under optimized conditions, the biosensor has a wide linear detection range of 0.1 pM to 1 μM, with a detection limit of 86.9 fM. Attribute to its high sensitivity, specificity, reproducibility and stability, this biosensor possesses considerable potential in early diagnosis of disease and biomedical research.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108844"},"PeriodicalIF":4.8,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142613379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-28DOI: 10.1016/j.bioelechem.2024.108842
Sri Ramulu Torati , Gymama Slaughter
C-reactive protein (CRP) is a critical biomarker for detecting inflammation and forecasting cardiovascular disease. We present an advanced electrochemical immunosensor utilizing laser-induced graphene (LIG)/MXene-gold nanoparticles (Mx-AuNPs) electrode for CRP detection. The Mx-AuNPs nanocomposite, synthesized via in-situ reduction of HAuCl4 by MXene, leverages MXene’s reducing properties for effective nanoparticle deposition, confirmed through scanning electron microscopy. This electrode demonstrates superior electrochemical performance due to enhanced surface area and synergy between LIG and Mx-AuNPs, improving overall electrode conductivity. The A-CRP antibody, immobilized via a cysteamine linker, enables CRP detection. The immunosensor achieves excellent detection across 10 pg mL−1 to 10 µg mL−1 CRP, with a low detection limit of 1.45 pg mL−1, and shows high selectivity for CRP. This LIG/Mx-AuNPs-based immunosensor is promising for sensitive CRP detection, aiding early cardiovascular disease diagnosis and improving patient outcomes.
{"title":"Advanced laser-induced graphene-based electrochemical immunosensor for the detection of C-reactive protein","authors":"Sri Ramulu Torati , Gymama Slaughter","doi":"10.1016/j.bioelechem.2024.108842","DOIUrl":"10.1016/j.bioelechem.2024.108842","url":null,"abstract":"<div><div>C-reactive protein (CRP) is a critical biomarker for detecting inflammation and forecasting cardiovascular disease. We present an advanced electrochemical immunosensor utilizing laser-induced graphene (LIG)/MXene-gold nanoparticles (Mx-AuNPs) electrode for CRP detection. The Mx-AuNPs nanocomposite, synthesized via in-situ reduction of HAuCl<sub>4</sub> by MXene, leverages MXene’s reducing properties for effective nanoparticle deposition, confirmed through scanning electron microscopy. This electrode demonstrates superior electrochemical performance due to enhanced surface area and synergy between LIG and Mx-AuNPs, improving overall electrode conductivity. The A-CRP antibody, immobilized via a cysteamine linker, enables CRP detection. The immunosensor achieves excellent detection across 10 pg mL<sup>−1</sup> to 10 µg mL<sup>−1</sup> CRP, with a low detection limit of 1.45 pg mL<sup>−1</sup>, and shows high selectivity for CRP. This LIG/Mx-AuNPs-based immunosensor is promising for sensitive CRP detection, aiding early cardiovascular disease diagnosis and improving patient outcomes.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108842"},"PeriodicalIF":4.8,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563475","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-28DOI: 10.1016/j.bioelechem.2024.108840
Zixi Li , Xinyue Su , Yihong Lin, Yu Zhang, Anlan Zhang, Xin Wu, Xi Jiyu, Qin Li, Zewen Wei
CRISPR/Cas9-mediated gene editing offers promising and safe therapeutic options for a wide range of diseases. The technical difficulty of efficiently acquiring large quantities of gene-edited therapeutic cells in a short time period is now preventing the widespread clinical application of CRISPR/Cas9-mediated gene editing. Herein, a Large Volume Continuous Electroporation Chip (LaViE-Chip) has been developed to address the challenge of acquiring sufficient quantities of genetically edited cells for CRISPR/Cas9 gene editing. By connecting multiple relatively narrow microfluidic channels in parallel, a satisfactory balance between cell flow volume and electric field uniformity was achieved with two simple off-chip electrodes, which also isolated harmful effects around electrodes from target cells. Meanwhile, by carefully designing the curvature of the microfluidic channel, hydrodynamic controlled rotation of target cells has been realized to improve the transfection efficiency and cell viability. With these improvements, the LaViE-Chip realized 71.06 % electrotransfection efficiency, 84.3 % cell viability, and 107 cell/min cell processing speed. Moreover, the first successful incessant CRISPR gene editing by electroporation has been demonstrated, laying the technical foundation of therapeutic CRISPR gene editing.
{"title":"Expanding the cell quantity of CRISPR/Cas9 gene editing by continuous microfluidic electroporation chip","authors":"Zixi Li , Xinyue Su , Yihong Lin, Yu Zhang, Anlan Zhang, Xin Wu, Xi Jiyu, Qin Li, Zewen Wei","doi":"10.1016/j.bioelechem.2024.108840","DOIUrl":"10.1016/j.bioelechem.2024.108840","url":null,"abstract":"<div><div>CRISPR/Cas9-mediated gene editing offers promising and safe therapeutic options for a wide range of diseases. The technical difficulty of efficiently acquiring large quantities of gene-edited therapeutic cells in a short time period is now preventing the widespread clinical application of CRISPR/Cas9-mediated gene editing. Herein, a Large Volume Continuous Electroporation Chip (LaViE-Chip) has been developed to address the challenge of acquiring sufficient quantities of genetically edited cells for CRISPR/Cas9 gene editing. By connecting multiple relatively narrow microfluidic channels in parallel, a satisfactory balance between cell flow volume and electric field uniformity was achieved with two simple off-chip electrodes, which also isolated harmful effects around electrodes from target cells. Meanwhile, by carefully designing the curvature of the microfluidic channel, hydrodynamic controlled rotation of target cells has been realized to improve the transfection efficiency and cell viability. With these improvements, the LaViE-Chip realized 71.06 % electrotransfection efficiency, 84.3 % cell viability, and 10<sup>7</sup> cell/min cell processing speed. Moreover, the first successful incessant CRISPR gene editing by electroporation has been demonstrated, laying the technical foundation of therapeutic CRISPR gene editing.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108840"},"PeriodicalIF":4.8,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-26DOI: 10.1016/j.bioelechem.2024.108843
Yige Li, Yingying Cheng, Haoyi Ren, Tiantian Ji, Zhengyi Zhao, Hongling Li, Chenglin Hong
In this study, we developed a new system that using zinc-based metal–organic frameworks NH2-Zn-PTC as the donor and ZiF-8@PDA as the acceptor to achieve highly sensitive detection of carcinoembryonic antigen (CEA), using the fundamentals of electrochemiluminescence resonance energy transfer (ECL-RET). Firstly, the aggregation-induced quenching effect (ACQ) was eliminated by the coordination of PTC in MOF and the ECL signal was improved. Secondly, the ECL signal was further amplified by using Au NPs and amino groups as co-reaction promoters to generate more SO4.−. In addition, the introduction of ZiF-8@PDA as an acceptor and NH2-Zn-PTC as a donor took advantage of the feature of partial overlap of the UV–vis absorption spectrum and ECL emission spectra between the two, thereby effectively initiating the ECL-RET behavior, which improved the detection sensitivity of the sensor. The prepared immunosensor showed good linearity in the concentration range of 10−4 to 80 ng/mL with a detection limit of 18.20 fg/mL. This makes it promising for clinical testing of tumor markers.
{"title":"A sensitive electrochemiluminescence immunosensor for CEA detection based on the ECL-RET between zinc-based metal–organic frameworks and ZiF-8@PDA","authors":"Yige Li, Yingying Cheng, Haoyi Ren, Tiantian Ji, Zhengyi Zhao, Hongling Li, Chenglin Hong","doi":"10.1016/j.bioelechem.2024.108843","DOIUrl":"10.1016/j.bioelechem.2024.108843","url":null,"abstract":"<div><div>In this study, we developed a new system that using zinc-based metal–organic frameworks NH<sub>2</sub>-Zn-PTC as the donor and ZiF-8@PDA as the acceptor to achieve highly sensitive detection of carcinoembryonic antigen (CEA), using the fundamentals of electrochemiluminescence resonance energy transfer (ECL-RET). Firstly, the aggregation-induced quenching effect (ACQ) was eliminated by the coordination of PTC in MOF and the ECL signal was improved. Secondly, the ECL signal was further amplified by using Au NPs and amino groups as co-reaction promoters to generate more SO<sub>4</sub><sup>.−</sup>. In addition, the introduction of ZiF-8@PDA as an acceptor and NH<sub>2</sub>-Zn-PTC as a donor took advantage of the feature of partial overlap of the UV–vis absorption spectrum and ECL emission spectra between the two, thereby effectively initiating the ECL-RET behavior, which improved the detection sensitivity of the sensor. The prepared immunosensor showed good linearity in the concentration range of 10<sup>−4</sup> to 80 ng/mL with a detection limit of 18.20 fg/mL. This makes it promising for clinical testing of tumor markers.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108843"},"PeriodicalIF":4.8,"publicationDate":"2024-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Monitoring of the levels of 5-hydroxyindole-3-acetic acid (5-HIAA) is of significant importance for diagnostics of carcinoid tumors. We propose simple catalytic electrochemical sensors for the determination of 5-HIAA in urine using laccase and its mimetics. Laccase-like nanozymes (LacNZs) were synthesized via a chemical reduction, and resulting PtMn and MnO2 nanoflowers (NFs) demonstrated laccase-like activity similar to the laccase from the Trametes zonata. In addition, these LacNZs showed enhanced stability under a wide range of pH (3.0–7.5), temperatures (4–70 °C), and ionic strengths (up to 500 mM NaCl). The developed PtMn NF/graphite electrode, similar to a laccase/graphite electrode, can detect 5-HIAA with a high sensitivity (25 000 ± 12 A·M−1·m−2 and 1900 ± 9 A·M−1·m−2, respectively) and have linear ranges of 0.3 – 15 μM and 2 – 50 μM. The sensors work at low working potentials with a detection limit of 0.16 and 1.4 μM, covering the normal and pathologic ranges of 5-HIAA (1 – 50 μM) content in urine. They have been successfully applied to 5-HIAA assay in urine samples of people with various diseases and revealed good recovery values and reproducibility. Additionally, the LacNZ-sensor has the best stability and can be used up to 20 days.
{"title":"Laccase mimetics as sensing elements for amperometric assay of 5-hydroxyindoleacetic acid in urine","authors":"Olha Demkiv , Wojciech Nogala , Nataliya Stasyuk , Marcin Holdynski , Nina Dimcheva , Taras Danysh , Monika Asztemborska , Mykhailo Gonchar","doi":"10.1016/j.bioelechem.2024.108839","DOIUrl":"10.1016/j.bioelechem.2024.108839","url":null,"abstract":"<div><div>Monitoring of the levels of 5-hydroxyindole-3-acetic acid (5-HIAA) is of significant importance for diagnostics of carcinoid tumors. We propose simple catalytic electrochemical sensors for the determination of 5-HIAA in urine using laccase and its mimetics. Laccase-like nanozymes (LacNZs) were synthesized <em>via</em> a chemical reduction, and resulting PtMn and MnO<sub>2</sub> nanoflowers (NFs) demonstrated laccase-like activity similar to the laccase from the <em>Trametes zonata</em>. In addition, these LacNZs showed enhanced stability under a wide range of pH (3.0–7.5), temperatures (4–70 °C), and ionic strengths (up to 500 mM NaCl). The developed PtMn NF/graphite electrode, similar to a laccase/graphite electrode, can detect 5-HIAA with a high sensitivity (25 000 ± 12 A·M<sup>−1</sup>·m<sup>−2</sup> and 1900 ± 9 A·M<sup>−1</sup>·m<sup>−2</sup>, respectively) and have linear ranges of 0.3 – 15 μM and 2 – 50 μM. The sensors work at low working potentials with a detection limit of 0.16 and 1.4 μM, covering the normal and pathologic ranges of 5-HIAA (1 – 50 μM) content in urine. They have been successfully applied to 5-HIAA assay in urine samples of people with various diseases and revealed good recovery values and reproducibility. Additionally, the LacNZ-sensor has the best stability and can be used up to 20 days.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108839"},"PeriodicalIF":4.8,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-18DOI: 10.1016/j.bioelechem.2024.108834
Marina Serin , Pınar Kara
Multiple sclerosis (MS) is a recurrent inflammatory, demyelinating disease of the white matter in central nervous system (CNS). The number of MS patients is increasing, but the diagnostic process is still quite difficult, costly and requires combination of several methods. Myelin basic protein (MBP) makes up to 30 % of the myelin in CNS. It is known that MBP is released into the cerebrospinal fluid (CSF) as MS bioindicator. Herein, myelin specific DNA aptamer earlier developed for possible therapeutic purposes and anti-MBP antibody were applied as bioreceptors for MBP recognition on the same nanomodified sensor surfaces and their performances were compared. Biosensors were developed by using graphene oxide (GO) nanoparticles integrated onto pencil graphite electrodes (PGE) and bioreceptor molecules immobilized to create a bioactive layer for MBP binding. The measurements were run with electrochemical impedance spectroscopy (EIS). Selectivity of the biosensors was evaluated using human serum albumin (HSA). After optimization of binding parameters, biosensors were validated in artificial CSF. It was shown that LJM-5708 based aptasensor had LOD 0.65 ng/mL that was comparable to immunosensor LOD (0.36 ng/mL) in artificial CSF and showed its applicability in the clinical concentration range between 1 and 128 ng/mL.
{"title":"Demyelination detection in CSF based on electrochemical monitoring of myelin basic protein in comparison between Apta vs. Immuno sensing strategies","authors":"Marina Serin , Pınar Kara","doi":"10.1016/j.bioelechem.2024.108834","DOIUrl":"10.1016/j.bioelechem.2024.108834","url":null,"abstract":"<div><div>Multiple sclerosis (MS) is a recurrent inflammatory, demyelinating disease of the white matter in central nervous system (CNS). The number of MS patients is increasing, but the diagnostic process is still quite difficult, costly and requires combination of several methods. Myelin basic protein (MBP) makes up to 30 % of the myelin in CNS. It is known that MBP is released into the cerebrospinal fluid (CSF) as MS bioindicator. Herein, myelin specific DNA aptamer earlier developed for possible therapeutic purposes and anti-MBP antibody were applied as bioreceptors for MBP recognition on the same nanomodified sensor surfaces and their performances were compared. Biosensors were developed by using graphene oxide (GO) nanoparticles integrated onto pencil graphite electrodes (PGE) and bioreceptor molecules immobilized to create a bioactive layer for MBP binding. The measurements were run with electrochemical impedance spectroscopy (EIS). Selectivity of the biosensors was evaluated using human serum albumin (HSA). After optimization of binding parameters, biosensors were validated in artificial CSF. It was shown that LJM-5708 based aptasensor had LOD 0.65 ng/mL that was comparable to immunosensor LOD (0.36 ng/mL) in artificial CSF and showed its applicability in the clinical concentration range between 1 and 128 ng/mL.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108834"},"PeriodicalIF":4.8,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142491853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-16DOI: 10.1016/j.bioelechem.2024.108837
Seham O. Alsulami , Tadeusz Malinski , Howard D. Dewald
The impact of four clinically significant genetic variants of endothelial nitric oxide synthase (eNOS) polymorphisms on the concentrations of nitric oxide [NO] and peroxynitrite [ONOO–] has been given scant consideration. This study utilized a [NO]/[ONOO–] ratio to determine the extent of endothelial dysfunction caused by these variations in the eNOS gene. The single nucleotide polymorphisms (T-786C, C-665T, and Glu298Asp) and a variable number of tandem repeats (intron 4 a/b/c) were genotyped in human umbilical vein endothelial cells (HUVEC), using sanger sequencing and DNA electrophoresis, respectively. Nanosensors were used to determine the maximal [NO] and [ONOO–], while traditional and low-temperature SDS-PAGE were used to evaluate the expression of eNOS and the eNOS dimer-to-monomer ratio, respectively. The study results indicate that the eNOS haplotype H3 (G T/C C 4a/c allele) may have a protective effect against cardiovascular disease (CVD) with the [NO]/[ONOO–] ratio higher than 2. However, the eNOS haplotypes H2 (G T/C C 4a/b) and H5 (T T/C C 4b) increase the susceptibility to CVD with [NO]/[ONOO–] ratio lower than 1. The results suggest that certain eNOS genetic variants may influence susceptibility to cardiovascular disease (CVD) while other variants may have a protective effect.
内皮一氧化氮合酶（eNOS）多态性的四种具有临床意义的基因变异对一氧化氮[NO]和过氧化亚硝酸盐[ONOO-]浓度的影响一直鲜有研究。本研究利用[NO]/[ONOO-]比值来确定这些 eNOS 基因变异引起的内皮功能障碍的程度。利用桑格测序法和 DNA 电泳法分别对人脐静脉内皮细胞（HUVEC）中的单核苷酸多态性（T-786C、C-665T 和 Glu298Asp）和可变串联重复序列（内含子 4 a/b/c）进行了基因分型。纳米传感器用于测定最大[NO]和[ONOO-]，传统SDS-PAGE和低温SDS-PAGE分别用于评估eNOS的表达和eNOS二聚体与单体的比例。研究结果表明，当[NO]/[ONOO-]比值高于2时，eNOS单倍型H3（G T/C C 4a/c等位基因）可能对心血管疾病（CVD）有保护作用。研究结果表明，某些 eNOS 单倍型 H2（G T/C C 4a/b）和 H5（T T/C C 4b）可能会增加对[NO]/[ONOO-]比值低于 1 的心血管疾病的易感性。
{"title":"Influence of endothelial nitric oxide synthase haplotypes on nitric oxide and peroxynitrite productions","authors":"Seham O. Alsulami , Tadeusz Malinski , Howard D. Dewald","doi":"10.1016/j.bioelechem.2024.108837","DOIUrl":"10.1016/j.bioelechem.2024.108837","url":null,"abstract":"<div><div>The impact of four clinically significant genetic variants of endothelial nitric oxide synthase (eNOS) polymorphisms on the concentrations of nitric oxide [NO] and peroxynitrite [ONOO<sup>–</sup>] has been given scant consideration. This study utilized a [NO]/[ONOO<sup>–</sup>] ratio to determine the extent of endothelial dysfunction caused by these variations in the eNOS gene. The single nucleotide polymorphisms (T-786C, C-665T, and Glu298Asp) and a variable number of tandem repeats (intron 4 a/b/c) were genotyped in human umbilical vein endothelial cells (HUVEC), using sanger sequencing and DNA electrophoresis, respectively. Nanosensors were used to determine the maximal [NO] and [ONOO<sup>–</sup>], while traditional and low-temperature SDS-PAGE were used to evaluate the expression of eNOS and the eNOS dimer-to-monomer ratio, respectively. The study results indicate that the eNOS haplotype H3 (G T/C C 4a/c allele) may have a protective effect against cardiovascular disease (CVD) with the [NO]/[ONOO<sup>–</sup>] ratio higher than 2. However, the eNOS haplotypes H2 (G T/C C 4a/b) and H5 (T T/C C 4b) increase the susceptibility to CVD with [NO]/[ONOO<sup>–</sup>] ratio lower than 1. The results suggest that certain eNOS genetic variants may influence susceptibility to cardiovascular disease (CVD) while other variants may have a protective effect.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108837"},"PeriodicalIF":4.8,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142454374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-16DOI: 10.1016/j.bioelechem.2024.108838
Miao Wang , He-Nan Sun , Xing-Yu Liu , Mingjun Liu , Shan-Shan Li
Cardiac troponin I (cTnI) has been widely used in clinical diagnosis of acute myocardial infarction (AMI). Herein, a sensitive electrochemical biosensor for cTnI analysis was designed, in which the simple synthesized Pd@PdPtCo mesoporous nanopolyhedras (MNPs) were utilized as signal amplifiers. The mesoporous polyhedral structure of Pd@PdPtCo MNPs endows them with more specific surface area and more active sites, as well as the synergistic effect between multiple metal elements, all of which increase the electrocatalytic performance of Pd@PdPtCo MNPs in efficiently oxidizing hydroquinone (HQ) to benzoquinone (BQ). Experimental results showed that Pd@PdPtCo MNPs had better performance in oxidation of HQ to BQ compared with their corresponding monometallic and bimetallic nanomaterials. With the aid of the interaction between antigens and antibodies, the peak current of HQ to BQ showed an upward trend with increasing concentration of cTnI, thus the quantitative detection of cTnI could be achieved. Under optimal conditions, the biosensor prepared in this work has a wider linear range (1.0 × 10−4–200 ng mL−1) and a lower detection limit (0.031 pg mL−1) than other sensors reported in literatures, coupled by good stability and high sensitivity. More importantly, it also performed well in complex serum environment, proving that the electrochemical sensor has a practical application potential in this field.
{"title":"A sensitive electrochemical biosensor based on Pd@PdPtCo mesoporous nanopolyhedras as signal amplifiers for assay of cardiac troponin I","authors":"Miao Wang , He-Nan Sun , Xing-Yu Liu , Mingjun Liu , Shan-Shan Li","doi":"10.1016/j.bioelechem.2024.108838","DOIUrl":"10.1016/j.bioelechem.2024.108838","url":null,"abstract":"<div><div>Cardiac troponin I (cTnI) has been widely used in clinical diagnosis of acute myocardial infarction (AMI). Herein, a sensitive electrochemical biosensor for cTnI analysis was designed, in which the simple synthesized Pd@PdPtCo mesoporous nanopolyhedras (MNPs) were utilized as signal amplifiers. The mesoporous polyhedral structure of Pd@PdPtCo MNPs endows them with more specific surface area and more active sites, as well as the synergistic effect between multiple metal elements, all of which increase the electrocatalytic performance of Pd@PdPtCo MNPs in efficiently oxidizing hydroquinone (HQ) to benzoquinone (BQ). Experimental results showed that Pd@PdPtCo MNPs had better performance in oxidation of HQ to BQ compared with their corresponding monometallic and bimetallic nanomaterials. With the aid of the interaction between antigens and antibodies, the peak current of HQ to BQ showed an upward trend with increasing concentration of cTnI, thus the quantitative detection of cTnI could be achieved. Under optimal conditions, the biosensor prepared in this work has a wider linear range (1.0 × 10<sup>−4</sup>–200 ng mL<sup>−1</sup>) and a lower detection limit (0.031 pg mL<sup>−1</sup>) than other sensors reported in literatures, coupled by good stability and high sensitivity. More importantly, it also performed well in complex serum environment, proving that the electrochemical sensor has a practical application potential in this field.</div></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":"161 ","pages":"Article 108838"},"PeriodicalIF":4.8,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142491852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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