Analytical biochemistry最新文献

{"title":"Esterase-2 mutant-based nanostructured amperometric biosensors for the selective determination of paraoxon (Neurotoxin).","authors":"Hend Samy Magar, Muhammad Fayez, Ferdinando Febbraio, Rabeay Y A Hassan","doi":"10.1016/j.ab.2024.115751","DOIUrl":"10.1016/j.ab.2024.115751","url":null,"abstract":"<p><p>Organophosphate pesticides (OPs) are causing non-selective inhibition in enzymatic bioreceptors, thus the enzymatic-inhibition-based traditional assays are not suitable for their specific detection in food and environmental samples. Accordingly, a selective nanostructured electrochemical biosensing system was designed using six mutants of the esterase-2 (EST2 protein) enzymes from A. acidocaldarius to be exploited as targeting bio-receptors for the specific detection of OPs. Each of the EST2 mutant enzymes was immobilized on disposable screen-printed electrodes modified with Aluminum oxide (Al₂O₃)/Copper (Cu) nanocomposite. Consequently, chronoamperometric assay was fully optimized, and cross-reactivity study was carried out using paraoxon, malathion and chlorpyrifos. The comparative cross-reactivity study was performed on the six mutant proteins in terms of inhibitory percentage over a wide range of pesticide concentrations. Eventually, a wide dynamic inhibition range was achieved while the limit of detection for the paraoxon toxicity was 0.01 nM and the limit of quantification was 0.05 nM. Finally, paraoxon was selectively determined using the newly developed EST-based biosensor in different spiked food samples.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115751"},"PeriodicalIF":2.6,"publicationDate":"2024-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142833303","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}

{"title":"Norovirus detection technologies: From conventional methods to innovative biosensors.","authors":"David Septian Sumanto Marpaung, Ayu Oshin Yap Sinaga, Damayanti Damayanti","doi":"10.1016/j.ab.2024.115750","DOIUrl":"10.1016/j.ab.2024.115750","url":null,"abstract":"<p><p>The norovirus (NoV), known for its high contagion rate, is the leading cause of acute gastroenteritis. The development of a NoV vaccine is hindered by significant antigenic variation, lack of suitable models, unknown vaccine protection duration, limited human challenge studies, complex performance patterns, and the absence of a reliable in vitro cultivation system, making prevention, early detection, and control the only effective measures to mitigate outbreaks. This review aims to discuss about several norovirus biosensor for point-of-care analysis. Several innovative biosensors have been developed, including techniques such as electrochemical NoV biosensors, colorimetric NoV biosensors, fluorescence NoV biosensors, CRISPR-based NoV biosensors, and other NoV biosensors. These approaches have detected norovirus in biological samples with high sensitivity and specificity. This biosensing technique holds significant promise, not only in improving the speed and accuracy of diagnostic processes but also in strengthening the global response to norovirus infections, thereby underscoring its pivotal role in public health and disease prevention.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115750"},"PeriodicalIF":2.6,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142823773","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}

{"title":"Comparative lipidomics profiles of planktonic and biofilms of methicillin-resistant and -susceptible Staphylococcus aureus.","authors":"Shilpa Saseendran Nair, Torsten Kleffmann, Briana Smith, Vanessa Morris, Christoph Göbl, Daniel Pletzer, Matthias Fellner","doi":"10.1016/j.ab.2024.115746","DOIUrl":"10.1016/j.ab.2024.115746","url":null,"abstract":"<p><p>Staphylococcus aureus is a significant human pathogen causing acute life-threatening, and chronic infections often linked to biofilms. This study conducted a comparative lipidomic analysis of a methicillin-resistant (MRSA) and a methicillin-susceptible (MSSA) S. aureus strain in both planktonic and biofilm cultures using liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) spectroscopy. The developed protocol successfully differentiates between the strains in various living states (planktonic and biofilm) and growth media (Tryptic Soy Broth and Brain Heart Infusion) using S. aureus USA300 LAC (MRSA) and S. aureus Newman (MSSA). LC-MS and NMR lipidomics profiles revealed global differences and particular ones among the following classes of bacterial lipids: phosphatidylglycerols, diacylglycerols, monoglycosyldiacylglycerols, diglycosyldiacylglycerols, and cardiolipins. Lipid content was higher in the biofilm states for most of these classes. Growth media differences were significant, while differences between MRSA and MSSA were less pronounced but still detectable. Additionally, we provide data on hundreds of unknown compounds that differ based on living state, strain background, or growth media. This study offer insights into the dynamic nature of S. aureus lipid composition and the used methods are adaptable to other organisms.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115746"},"PeriodicalIF":2.6,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142821694","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}

{"title":"Comparison of the techniques for isolating immunoassay-suitable proteins from heterogeneous fecal samples.","authors":"Subramaniam-Betty Sheila Devan, Rosli Ramli, Salah Abdalrazak Alshehade, Sharoen Yu Ming Lim, Noorhidayah Mamat","doi":"10.1016/j.ab.2024.115748","DOIUrl":"10.1016/j.ab.2024.115748","url":null,"abstract":"<p><p>Immunoassays could provide valuable insights into disease biomarkers and gut health by measuring fecal proteins. However, reliably isolating intact proteins from feces is challenging due to its heterogeneous and variable composition. This paper aims to review and compare different methods for extracting proteins from fecal samples to make them suitable for immunoassay analysis. Mechanical homogenization helps release proteins by disrupting solids, while protease inhibitors preserve protein integrity. Detergents like SDS solubilize proteins by disrupting hydrophobic interactions. Organic solvents such as acetone precipitate proteins and remove contaminants. Thermal treatment denatures proteases. Immunocapture uses antibodies to purify target proteins away from interference selectively. Commercial kits contain optimized buffers but may be cost-prohibitive. Combining mechanical, chemical, and immunological techniques synergistically integrates their advantages, improving the recovery of native proteins with reduced matrix effects. While all methods have merits, tailored protocols integrating multiple mechanisms appear most promising for extracting immunoassay-suitable fecal proteins. Further optimization and standardization of such combination approaches matched to proteins and assays of interest helps expand noninvasive fecal proteome analysis.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115748"},"PeriodicalIF":2.6,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142817003","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}

{"title":"Evaluation of active substances in gamboge and their mechanisms for the treatment of colorectal cancer by UPLC-MS/MS integrated with network pharmacology.","authors":"Guodong Shan, Jiajun Jiang, Liting Ji, Shiyan Li, Zejun Wang, Shaohui Yang, Qing Shen","doi":"10.1016/j.ab.2024.115747","DOIUrl":"10.1016/j.ab.2024.115747","url":null,"abstract":"<p><p>Gamboge exhibits anti-colorectal cancer (CRC) activity, however, its active compounds and the underlying mechanisms remain unclear. Herein, a liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for determining gambogellic acid, β-morellic acid, isogambogenic acid, gambogenic acid, R-gambogic acid, S-gambogic acid, and hydroxygambogic acid in gamboge was established. The key parameters including ion transitions, voltages, LOD, and LOQ were determined, with LOD ranging from 0.8 to 2.0 ng mL^-1 and LOQ from 2.7 to 6.7 ng mL^-1. The recovery rates were found to be between 95.6 % and 103.5 %. Furthermore, the active compounds were successfully determined, and molecular mechanisms of gamboge in treating CRC were explored. Network pharmacology revealed a \"compound-target-pathway\" network where the seven compounds could target key proteins, modulate PI3K-Akt and JAK-STAT pathways, and inhibit CRC development. Molecular docking validated SRC, SATA3, PIK3CA, among others, as potential targets for the active compounds in CRC intervention. In conclusion, this method significantly reduces analysis time and improves efficiency relative to existing approaches, making it highly suitable for the effective determination of multiple compounds in the quality control of gamboge materials.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115747"},"PeriodicalIF":2.6,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794247","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}

{"title":"The application of different biotechnologies in detecting the changes in MAM and their classic discoveries.","authors":"Shuangshuang Zhang, Yingchun Shao, Mengzhu Su, Yuerong Hao, Yang Yuan, Dongming Xing","doi":"10.1016/j.ab.2024.115744","DOIUrl":"10.1016/j.ab.2024.115744","url":null,"abstract":"<p><p>Mitochondria-associated membrane (MAM) has been studied as a novel target for explaining the mechanisms underlying the changes in cellular function and the process of multiple diseases. This structure is a complex of proteins, it tethers mitochondria to the endoplasmic/sarcoplasmic reticulum (ER/SR) and mediates the crosstalk of ions, lipids and metabolites between the two organelles. Different component proteins play distinctive ways in influencing the structure of MAM or the cellular signal transduction. Mitochondria and ER are the hubs of cellular bioenergetics and protein homeostasis respectively, MAM was supposed to play both physiological and pathological roles in regulating the function of either the two organelles and cells. The mitochondria-associated membrane is a highly dynamic structure and could be disrupted or remodelled within several minutes. Up to now, not all component proteins of the MAM complex have been revealed. Several biochemical and imaging approaches have been used to measure the changes in the structure or the number of MAMs, but they come with their issues. For distinct research aims, particular techniques were used based on their applicabilities, the research platforms and technical defects. This review briefly summarized the current biotechnologies for detecting MAM, and analyzed their limitations, aiming to assist further research in selecting appropriate methods based on actual situations.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115744"},"PeriodicalIF":2.6,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794248","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}

{"title":"Cryopreserved PBMCs can be used for the analysis of mitochondrial respiration and serve as a diagnostic tool for mitochondrial diseases.","authors":"Zuzana Korandová, Petr Pecina, Alena Pecinová, Eliška Koňaříková, Markéta Tesařová, Josef Houštěk, Hana Hansíková, Hana Ptáčková, Jiří Zeman, Tomáš Honzík, Tomáš Mráček","doi":"10.1016/j.ab.2024.115745","DOIUrl":"10.1016/j.ab.2024.115745","url":null,"abstract":"<p><p>Mitochondrial diseases are severe, inherited metabolic disorders that affect the paediatric population. They affect the functioning of mitochondrial oxidative phosphorylation (OXPHOS) apparatus either directly or indirectly. Since mutations in mtDNA are responsible for only 25 % of paediatric cases and next-generation sequencing does not always provide a conclusive diagnosis, the biochemical approach still represents a valuable tool in diagnostics. Mitochondrial defects can be identified in tissue biopsies (muscle or skin). However, they also often manifest in peripheral blood cells. We developed a protocol for isolation and cryopreservation of peripheral blood mononuclear cells (PBMCs) from 5 ml of children's blood using Ficoll centrifugation which can be utilised for subsequent functional measurements on thawed samples. Furthermore, we evaluated the diagnostic utility of the optimised high-resolution oxygraphy protocol using digitonin-permeabilized cryopreserved PBMCs on 47 samples from patients with confirmed or suspected mitochondrial disease. Overall, the diagnosis was confirmed in 72 % of cases, while the analysis of cryopreserved PBMCs provided a false negative outcome in 13 % of cases. Our study demonstrates a sensitive, fast, and non-invasive approach for the diagnostics of various types of mitochondrial disorders, especially those of nuclear genetic origin manifesting in paediatric patients.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115745"},"PeriodicalIF":2.6,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791002","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}

{"title":"Electrochemical determination of ascorbic acid using sensitive and disposable methylene blue modified pencil graphite electrode.","authors":"K G Aishwarya, Y Arthoba Nayaka, E Pradeepa, H R Sahana","doi":"10.1016/j.ab.2024.115733","DOIUrl":"10.1016/j.ab.2024.115733","url":null,"abstract":"<p><p>In the present work, a convenient, efficient and disposable electrochemical sensor has been developed by electropolymerizing methylene blue (PMB) on the surface of a pencil graphite electrode (PGE), which facilitates the electrochemical analysis of an antioxidant l-Ascorbic Acid (AA). The structural characteristics of both the methylene blue modified pencil graphite electrode (PMB/PGE) and the bare pencil graphite electrode (BPGE) have been examined using scanning electron microscopy (SEM) in conjunction with energy-dispersive X-ray analysis (EDX). Additionally, the charge transfer behavior has been evaluated using the electron impedance spectroscopy (EIS). The voltammetric response of AA has been examined using different methods, such as differential pulse voltammetry (DPV) and linear sweep voltammetry (LSV). This exploration has been carried out in 0.1 M phosphate buffer solution (PBS) of physiological pH 7.0. The electrochemical sensor PMB/PGE proposed in this study exhibited an improved peak current and a slight negative shift in peak potential for AA compared to bare electrode. The enhancement in peak current at the modified electrode has been attributed to the electrocatalytic characteristics of the modifiers. The limit of detection (LOD) for AA has been determined using the differential pulse voltammetry (DPV), with concentrations ranging from 1.0 μM to 12.0 μM. The calculated LOD value has been found to be 0.15 μM. The selectivity and practicality of the modified electrode has been assessed through the real sample analysis and demonstrating its capability to detect AA in the presence of paracetamol (PA) resulting in satisfactory recovery results. Hence the proposed sensor could be successfully validated for the determination of AA in pharmaceutical sample.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115733"},"PeriodicalIF":2.6,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142779444","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}

{"title":"Development and validation of a sensitive and fast bioanalytical LC-MS/MS assay for the quantitation of venetoclax and azacitidine in Rat Plasma: Application to pharmacokinetic study.","authors":"Manal El-Gendy, Mohamed Hefnawy, Sarah Alrubia, Abdulaziz Alnasser, Amsha Alsegiani, Yousef Bin Jardan, Adel El-Azab, Alaa Abdel-Aziz, Mohamed Attwa, Emad Alsarhani","doi":"10.1016/j.ab.2024.115741","DOIUrl":"10.1016/j.ab.2024.115741","url":null,"abstract":"<p><p>The combination of venetoclax plus azacitidine (VTX-AZA) is FDA-approved to treat patients with acute myeloid leukemia (AML) aged ≥75 years and has become the standard of care for AML patients. However, the literature has not reported an analytical method for determining VTX-AZA in plasma samples. Therefore, developing an accurate and sensitive bioanalytical assay to quantify VTX-AZA in plasma is important. For the first time, this study describes the development of a new liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for the simultaneous determination of VTX and AZC in plasma samples with its application to pharmacokinetic study in rats. The assay employs repaglinide (RPG) as an internal standard. The chromatographic separations of VTX, AZC, and RPG are achieved within 2.5 min at 25 °C on an Eclipse plus C18 column (100 mm × 2.1 mm, 1.8 μm) and an isocratic mobile phase consisted of water with 0.1 % formic acid and acetonitrile (50:50, v/v, pH 3.2) at a flow rate of 0.30 mL/min. VTX and AZC have been extracted from rat plasma using the solid-phase extraction (SPE) procedure without interference from plasma endogenous. The FDA guidelines were followed in the validation of the developed assay, and linearity in rat plasma was observed for AZC and VTX, respectively, ranging from 5 to 3000 and 5-1000 ng/mL, with r ≥ 0.998. The lower limits of detection (LLOD) were 2 ng/mL for both drugs. In addition, the inter-day and intra-day accuracy were 0.8-6.6 % and 2.2-5.7 %; the inter-day and intra-day precision were 3-6.6 % and 1.5-7.1 %, respectively. The validated assay was effectively used in a pharmacokinetic investigation including the simultaneous oral administration of 40 mg/kg of AZA and 100 mg/kg of VTX to rats. The maximum plasma concentration (C_max) for AZC and VTX was 794 ± 99.6 ng/mL and 641 ± 96.9 ng/mL achieved at 0.5 ± 0.03 h and 6 ± 0.05 h, respectively. The AUC_0-∞ for AZC and VTX was 1253 ± 252.6 and 4881 ± 745.4 ng/mL.h; respectively.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115741"},"PeriodicalIF":2.6,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142783862","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}

{"title":"High-resolution targeted mass spectrometry for comprehensive quantification of sphingolipids: clinical applications and characterization of extracellular vesicles.","authors":"Thiago V D Felippe, Diana M Toro, Jonatan C S de Carvalho, Pedro Nobre-Azevedo, Luiz F M Rodrigues, Bianca T M Oliveira, Pedro V da Silva-Neto, Adriana F L Vilela, Fausto Almeida, Lúcia H Faccioli, Carlos A Sorgi","doi":"10.1016/j.ab.2024.115732","DOIUrl":"10.1016/j.ab.2024.115732","url":null,"abstract":"<p><p>Sphingolipids (SL), a class of membrane lipids, play important roles in numerous biological processes. Their significant structural diversity poses challenges for accurate quantification. To address this, liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has emerged as a powerful tool for sphingolipidomics, capable of profiling these lipids comprehensively. In this study, we utilized LC-MS/MS with high-resolution mass spectrometry (MRM^HR) to develop a targeted method for the identification and quantification of various SL species. This method, based on validated parameters such as precursor/fragment ions (m/z) and retention time, demonstrated high sensitivity and accuracy, successfully identifying SL species across 12 distinct classes. Its open-panel design also facilitates the analysis of new SL-species targets. Notably, using this approach, we identified 40 SL species in plasma samples from COVID-19 patients, and we determined the influence of matrix metalloproteinase-3 (MMP-3) expression on the positive downstream of SL metabolism. Beyond plasma analysis, this method has potential applications in other biomedical contexts, such as extracellular vesicles (EVs), describing the cargo of sphingosine-1-phosphate (S1P) on macrophage-derived EVs. The establishment of this targeted workflow enabling precise quantification of a wide range of SL species, holds promise for identifying novel biomarkers and therapeutic targets.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115732"},"PeriodicalIF":2.6,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765574","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}