{"title":"利用表面工程 AuPt@GQD 核壳纳米复合材料制备无标记免疫传感器,用于选择性检测受污染食品样品中的痕量大肠埃希氏菌。","authors":"Tushar Das, Subrata Das* and Betty C. A*, ","doi":"10.1021/acsbiomaterials.4c00297","DOIUrl":null,"url":null,"abstract":"<p >Fabrication of label-free immunosensors is highly necessitated due to their simplicity, cost-effectiveness, and robustness. Herein, we report the facile development of a label-free, direct, rapid, capacitive immunosensor for ultrasensitive and rapid recognition of trace levels of <i>Escherichia coli</i> from contaminated food samples. This was achieved using gold platinum core–shell nanoparticles loaded with graphene quantum dots (AuPt@GQDs) that were utilized as electrode modifiers. The incorporation of GQDs to the surface of AuPt core–shell nanoparticles was performed using the “<i>greener</i>” probe-sonication method. The electrochemical properties of AuPt@GQDs, determined using cyclic voltammetry and electrochemical impedance spectroscopy, suggested the optimized loading concentration of AuPt to be 0.05% in the core–shell nanocomposite to exhibit the highest current response. Furthermore, immobilization of anti-<i>E. coli</i> monoclonal antibodies (anti-<i>E. coli</i> mAb) onto the surface of modified electrodes was performed using amine coupling. The high specific binding of <i>E. coli</i> cells onto the surface of the immuno-electrode was measured as a direct function of change in transient capacitance with time that was measured at low and high frequencies. The resultant immunosensor (bovine serum albumin/anti-<i>E. coli</i> mAb/AuPt<sub>0.05</sub>@GQDs/FTO) demonstrated a detection range (5 to 4.5 × 10<sup>3</sup> cells/mL), with the detection limit as low as 1.5 × 10<sup>2</sup> cells/mL, and an excellent sensitivity ∼171,281.40 μF<sup>–1</sup> mL cells<sup>–1</sup> cm<sup>–2</sup> without the use of any labels (<i>R</i><sup>2</sup>-0.99). These findings were further verified using real sample analysis wherein the immuno-electrode demonstrated outstanding sensitivity, the highest noticed so far. More interestingly, the high resuability ∼48 weeks (RSD-5.92%) and excellent reproducibility in detection results (RSD ∼ 9.5%) testify its potential use in a clinical setting. The results reveal the usefulness of the surface-engineered AuPt@GQDs core–shell nanocomposite as an electrode modifier that can be used for the development of newer on-site monitoring devices to estimate trace levels of pathogens present as contaminants in food samples.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":null,"pages":null},"PeriodicalIF":5.4000,"publicationDate":"2024-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Fabrication of a Label-Free Immunosensor Using Surface-Engineered AuPt@GQD Core–Shell Nanocomposite for the Selective Detection of Trace Levels of Escherichia coli from Contaminated Food Samples\",\"authors\":\"Tushar Das, Subrata Das* and Betty C. A*, \",\"doi\":\"10.1021/acsbiomaterials.4c00297\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >Fabrication of label-free immunosensors is highly necessitated due to their simplicity, cost-effectiveness, and robustness. Herein, we report the facile development of a label-free, direct, rapid, capacitive immunosensor for ultrasensitive and rapid recognition of trace levels of <i>Escherichia coli</i> from contaminated food samples. This was achieved using gold platinum core–shell nanoparticles loaded with graphene quantum dots (AuPt@GQDs) that were utilized as electrode modifiers. The incorporation of GQDs to the surface of AuPt core–shell nanoparticles was performed using the “<i>greener</i>” probe-sonication method. The electrochemical properties of AuPt@GQDs, determined using cyclic voltammetry and electrochemical impedance spectroscopy, suggested the optimized loading concentration of AuPt to be 0.05% in the core–shell nanocomposite to exhibit the highest current response. Furthermore, immobilization of anti-<i>E. coli</i> monoclonal antibodies (anti-<i>E. coli</i> mAb) onto the surface of modified electrodes was performed using amine coupling. The high specific binding of <i>E. coli</i> cells onto the surface of the immuno-electrode was measured as a direct function of change in transient capacitance with time that was measured at low and high frequencies. The resultant immunosensor (bovine serum albumin/anti-<i>E. coli</i> mAb/AuPt<sub>0.05</sub>@GQDs/FTO) demonstrated a detection range (5 to 4.5 × 10<sup>3</sup> cells/mL), with the detection limit as low as 1.5 × 10<sup>2</sup> cells/mL, and an excellent sensitivity ∼171,281.40 μF<sup>–1</sup> mL cells<sup>–1</sup> cm<sup>–2</sup> without the use of any labels (<i>R</i><sup>2</sup>-0.99). These findings were further verified using real sample analysis wherein the immuno-electrode demonstrated outstanding sensitivity, the highest noticed so far. More interestingly, the high resuability ∼48 weeks (RSD-5.92%) and excellent reproducibility in detection results (RSD ∼ 9.5%) testify its potential use in a clinical setting. The results reveal the usefulness of the surface-engineered AuPt@GQDs core–shell nanocomposite as an electrode modifier that can be used for the development of newer on-site monitoring devices to estimate trace levels of pathogens present as contaminants in food samples.</p>\",\"PeriodicalId\":8,\"journal\":{\"name\":\"ACS Biomaterials Science & Engineering\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2024-05-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Biomaterials Science & Engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://pubs.acs.org/doi/10.1021/acsbiomaterials.4c00297\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MATERIALS SCIENCE, BIOMATERIALS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Biomaterials Science & Engineering","FirstCategoryId":"5","ListUrlMain":"https://pubs.acs.org/doi/10.1021/acsbiomaterials.4c00297","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
Fabrication of a Label-Free Immunosensor Using Surface-Engineered AuPt@GQD Core–Shell Nanocomposite for the Selective Detection of Trace Levels of Escherichia coli from Contaminated Food Samples
Fabrication of label-free immunosensors is highly necessitated due to their simplicity, cost-effectiveness, and robustness. Herein, we report the facile development of a label-free, direct, rapid, capacitive immunosensor for ultrasensitive and rapid recognition of trace levels of Escherichia coli from contaminated food samples. This was achieved using gold platinum core–shell nanoparticles loaded with graphene quantum dots (AuPt@GQDs) that were utilized as electrode modifiers. The incorporation of GQDs to the surface of AuPt core–shell nanoparticles was performed using the “greener” probe-sonication method. The electrochemical properties of AuPt@GQDs, determined using cyclic voltammetry and electrochemical impedance spectroscopy, suggested the optimized loading concentration of AuPt to be 0.05% in the core–shell nanocomposite to exhibit the highest current response. Furthermore, immobilization of anti-E. coli monoclonal antibodies (anti-E. coli mAb) onto the surface of modified electrodes was performed using amine coupling. The high specific binding of E. coli cells onto the surface of the immuno-electrode was measured as a direct function of change in transient capacitance with time that was measured at low and high frequencies. The resultant immunosensor (bovine serum albumin/anti-E. coli mAb/AuPt0.05@GQDs/FTO) demonstrated a detection range (5 to 4.5 × 103 cells/mL), with the detection limit as low as 1.5 × 102 cells/mL, and an excellent sensitivity ∼171,281.40 μF–1 mL cells–1 cm–2 without the use of any labels (R2-0.99). These findings were further verified using real sample analysis wherein the immuno-electrode demonstrated outstanding sensitivity, the highest noticed so far. More interestingly, the high resuability ∼48 weeks (RSD-5.92%) and excellent reproducibility in detection results (RSD ∼ 9.5%) testify its potential use in a clinical setting. The results reveal the usefulness of the surface-engineered AuPt@GQDs core–shell nanocomposite as an electrode modifier that can be used for the development of newer on-site monitoring devices to estimate trace levels of pathogens present as contaminants in food samples.
期刊介绍:
ACS Biomaterials Science & Engineering is the leading journal in the field of biomaterials, serving as an international forum for publishing cutting-edge research and innovative ideas on a broad range of topics:
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Characterization, Synthesis, and Modification – new biomaterials, bioinspired and biomimetic approaches to biomaterials, exploiting structural hierarchy and architectural control, combinatorial strategies for biomaterials discovery, genetic biomaterials design, synthetic biology, new composite systems, bionics, polymer synthesis
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Modeling and Informatics Tools – scaling methods to guide biomaterial design, predictive algorithms for structure-function, biomechanics, integrating bioinformatics with biomaterials discovery, metabolomics in the context of biomaterials
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