Competitive immunosensor based on gliadin immobilization on disposable carbon-nanogold screen-printed electrodes for rapid determination of celiotoxic prolamins

Manfredi, Anita; Giannetto, Marco; Mattarozzi, Monica; Costantini, Monica; Mucchino, Claudio; Careri, Maria

doi:10.1007/s00216-016-9494-z

Cited by 29 publications

(17 citation statements)

References 24 publications

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“…It uses gold nanoparticles-SPCE and alkaline phosphatase as an enzyme. The limit of detection was 8 µg L -1 gliadin, more than an order of magnitude larger than the one provided with this aptasensor [10]. As mentioned above, the aptamer-based electrochemical magnetoassay has a reported limit of detection of 0.5 mg kg -1 .…”

Section: Resultsmentioning

confidence: 77%

“…This indicates that there is a long way to reliably measure gluten in food samples using immunochemical approaches. Only two electrochemical immunosensors have been reported and used in real samples: a sandwich immunosensor that needs multiple steps to covalently attach the capture antibody to a self-assembled monolayer in addition to the binding and labeling steps and a 10-min enzymatic reaction before measurement [9]; and a competitive immunosensor with a limit of detection above 100 mg kg -1 due to incompatibility with extraction reagents [10].…”

Section: Introductionmentioning

confidence: 99%

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Disposable electrochemical aptasensor for gluten determination in food

López-López

Miranda‐Castro

de‐los‐Santos‐Álvarez

et al. 2017

Sensors and Actuators B: Chemical

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Reliable detection of decreasing amounts of gluten in food is the only way of ensuring the safety of all coeliac patients. Results obtained with the method of choice, immunochemical assays, are not entirely comparable and many of them are sandwich assays that cannot recognize hydrolyzed proteins. In this work, we propose a competitive electrochemical sensor based on a recently described aptamer targeting the gliadin immunodominant peptide 33-mer that triggers the coeliac disease. The sensing layer is built on the surface of a screen-printed carbon electrode (SPCE) by adsorption of streptavidin and subsequent peptide immobilization. A competition between the peptide and gluten proteins from samples for a defined concentration of biotinylated Gli 4 aptamer is established. The aptamer bound to the peptide on the surface is finally measured after enzyme labelling and chronoamperometric detection of an enzymatically obtained electrochemically active product. This method is able to detect as low as 0.113 µg L-1 of gliadin, which corresponds to 380 µg kg-1 of gluten in food, taking all dilutions and conversion factors into consideration, with a reproducibility lower than 11%. The aptasensor was applied to food samples with gluten contents above and below the legislated threshold for gluten-free labelling in the EU, obtaining good agreement with the official R5 immunochemical method. mg kg -1Gluten Threshold gluten-free S P eaptasensor Graphical Abstract (for review) Highlights An electrochemical aptamer-based sensor on peptide-modified carbon screen-printed electrodes for gluten determination  Sensitivity is improved compared with reference enzyme-linked immunosorbent assays  The sensor was successfully applied to detect gluten in various food samples AbstractReliable detection of decreasing amounts of gluten in food is the only way of ensuring the safety of all coeliac patients. Results obtained with the method of choice, immunochemical assays, are not entirely comparable and many of them are sandwich assays that cannot recognize hydrolyzed proteins. In this work, we propose a competitive electrochemical sensor based on a recently described aptamer targeting the gliadin immunodominant peptide 33-mer that triggers the coeliac disease. The sensing layer is built on the surface of a screen-printed carbon electrode (SPCE) by adsorption of streptavidin and subsequent peptide immobilization. A competition between the peptide and gluten proteins from samples for a defined concentration of biotinylated Gli 4 aptamer is established. The aptamer bound to the peptide on the surface is finally measured after enzyme labelling and chronoamperometric detection of an enzymatically obtained electrochemically active product. This method is able to detect as low as 0.113 µg L -1 of gliadin, which corresponds to 380 µg kg -1 of gluten in food, taking all dilutions and conversion factors into consideration, with a reproducibility lower than 11%. The aptasensor was applied to food samples with gluten contents above and below the legislat...

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Section: Resultsmentioning

confidence: 77%

Section: Introductionmentioning

confidence: 99%

Disposable electrochemical aptasensor for gluten determination in food

López-López

Miranda‐Castro

de‐los‐Santos‐Álvarez

et al. 2017

Sensors and Actuators B: Chemical

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“…The microfluidic e-tongue capable of detecting as low as 0.005 ppm of gliadin in foodstuffs has been investigated (344). The competitive and disposable amperometric immunosensor based on gliadin-functionalized carbon/nanogold screen-printed electrodes was developed for rapid gluten detection in processed food samples (345). The gliadin-immunochips, based on electrochemical impedance spectroscopy transduction method, capable of detecting 0.5 ppm of gliadin in beers and flours have been reported (346).…”

Section: Methods For Detection Of Gluten In Foodmentioning

confidence: 99%

Pathogenesis of Celiac Disease and Other Gluten Related Disorders in Wheat and Strategies for Mitigating Them

et al. 2020

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Wheat is a major cereal crop providing energy and nutrients to the billions of people around the world. Gluten is a structural protein in wheat, that is necessary for its dough making properties, but it is responsible for imparting certain intolerances among some individuals, which are part of this review. Most important among these intolerances is celiac disease, that is gluten triggered T-cell mediated autoimmune enteropathy and results in villous atrophy, inflammation and damage to intestinal lining in genetically liable individuals containing human leukocyte antigen DQ2/DQ8 molecules on antigen presenting cells. Celiac disease occurs due to presence of celiac disease eliciting epitopes in gluten, particularly highly immunogenic alpha-gliadins. Another gluten related disorder is non-celiac gluten-sensitivity in which innate immune-response occurs in patients along with gastrointestinal and non-gastrointestinal symptoms, that disappear upon removal of gluten from the diet. In wheat allergy, either IgE or non-IgE mediated immune response occurs in individuals after inhalation or ingestion of wheat. Following a lifelong gluten-free diet by celiac disease and non-celiac gluten-sensitivity patients is very challenging as none of wheat cultivar or related species stands safe for consumption. Hence, different molecular biology, genetic engineering, breeding, microbial, enzymatic, and chemical strategies have been worked upon to reduce the celiac disease epitopes and the gluten content in wheat. Currently, only 8.4% of total population is affected by wheat-related issues, while rest of population remains safe and should not remove wheat from the diet, based on false media coverage.

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“…A competitive disposable amperometric immunosensor was firstly developed by Manfredi et al [41] based on gliadin-functionalized carbon/nanogold screen-printed electrodes for rapid determination of celiotoxic prolamins. The developed immunocompetitive assay achieved good sensitivity for gliadin in ethanol extracts, giving limit of detection and limit of quantitation at 8 and 22 ng·mL −1 , respectively.…”

Section: Electrochemical Immunosensors With Different Signalsmentioning

confidence: 99%

Nanomaterials for Electrochemical Immunosensing

Pan

Yun

et al. 2017

Sensors

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Electrochemical immunosensors resulting from a combination of the traditional immunoassay approach with modern biosensors and electrochemical analysis constitute a current research hotspot. They exhibit both the high selectivity characteristics of immunoassays and the high sensitivity of electrochemical analysis, along with other merits such as small volume, convenience, low cost, simple preparation, and real-time on-line detection, and have been widely used in the fields of environmental monitoring, medical clinical trials and food analysis. Notably, the rapid development of nanotechnology and the wide application of nanomaterials have provided new opportunities for the development of high-performance electrochemical immunosensors. Various nanomaterials with different properties can effectively solve issues such as the immobilization of biological recognition molecules, enrichment and concentration of trace analytes, and signal detection and amplification to further enhance the stability and sensitivity of the electrochemical immunoassay procedure. This review introduces the working principles and development of electrochemical immunosensors based on different signals, along with new achievements and progress related to electrochemical immunosensors in various fields. The importance of various types of nanomaterials for improving the performance of electrochemical immunosensor is also reviewed to provide a theoretical basis and guidance for the further development and application of nanomaterials in electrochemical immunosensors.

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Competitive immunosensor based on gliadin immobilization on disposable carbon-nanogold screen-printed electrodes for rapid determination of celiotoxic prolamins

Cited by 29 publications

References 24 publications

Disposable electrochemical aptasensor for gluten determination in food

Disposable electrochemical aptasensor for gluten determination in food

Pathogenesis of Celiac Disease and Other Gluten Related Disorders in Wheat and Strategies for Mitigating Them

Nanomaterials for Electrochemical Immunosensing

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