Study of a New Gliadin Capture Agent and Development of a Protein Microarray as a New Approach for Gliadin Detection

Cimaglia, Fabio; Potente, Giovanni; Chiesa, Maurizio; Mita, Giovanni; Bleve, Gianluca

doi:10.4172/jpb.1000326

Cited by 4 publications

(3 citation statements)

References 60 publications

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“…Microarrays are based on the immobilization technology of gliadin and chymotrypsin digested gliadin and fluorescent-labeled glutamine binding protein. Microarrays seemed promising but not yet tested (Cimaglia et al, 2014).…”

Section: Novel Methodsmentioning

confidence: 99%

Gluten Detection in Foods

Munshi

Deb

2021

Challenges and Potential Solutions in Gluten Free Product Development

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Celiac disease is one of the most continuous perpetual food prejudices prompted by ingestion of gluten protein from various foods such as wheat, barley, rye, oats, etc. The accessibility of the various analytical methods is extremely important to determine the presence of gluten in the food matrix to guarantee the affluence of gluten delicate people. Along with it in accordance to Codex, foods having below 20 mg gluten/kg can only be considered under a gluten-free label. This also sets standards for the analytical methods in gluten detection. The present chapter deals with the chemical constituents, toxicity, and tolerance limit of gluten protein along with the importance of gluten-free foods, gluten labeling, and risk management. However, the main objective is to discuss the various gluten detection methods, their applicability, challenges, and influencing factors. Removal of gluten is necessary to increase the availability of gluten-free food products which greatly depends on the selection and standard of the detection method. Thus, to ensure the consumption of gluten-free food products the detection method of gluten must be monitored carefully.

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Section: Novel Methodsmentioning

confidence: 99%

Gluten Detection in Foods

Munshi

Deb

2021

Challenges and Potential Solutions in Gluten Free Product Development

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“…This protein is successfully used to detect gluten-digested peptides with higher affinity than the intact gluten by fluorescence on a protein microarray. However, this strategy also relies on peptide detection similarly to MS proteomics [14].…”

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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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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“…This method proved to be suitable for quantifying gliadins in real samples with a LOD of 0.6 μg/mL. An additional approach has been proposed by Cimaglia et al 17 who developed a protein microarray functionalized by using a recombinant glutamine-binding protein (Gln BP) and 4F3 monoclonal antibodies. Their device showed a good response up to 5 ppm of gliadin, although its application as a biosensor requires additional improvements.…”

Section: ■ Introductionmentioning

confidence: 99%

Label-Free Detection of Gliadin in Food by Quartz Crystal Microbalance-Based Immunosensor

Funari

Terracciano

Ventura

et al. 2017

J. Agric. Food Chem.

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Gluten is a protein composite found in wheat and related grains including barley, rye, oat, and all their species and hybrids. Gluten matrix is a biomolecular network of gliadins and glutenins that contribute to the texture of pastries, breads, and pasta. Gliadins are mainly responsible for celiac disease, one of the most widespread food-related pathologies in Western world. In view of the importance of gliadin proteins, by combining the quartz crystal microbalance technology, a cheap and robust piezoelectric transducer, with the so-called photonic immobilization technique, an effective surface functionalization method that provides spatially oriented antibodies on gold substrates, we realized a sensitive and reliable biosensor for quantifying these analytes extracted from real samples in a very short time. The resulting immunosensor has a limit of detection of about 4 ppm and, more remarkably, shows excellent sensitivity in the range 7.5-15 ppm. This feature makes our device reliable and effective for practical applications since it is able to keep low the influence of false positives.

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Study of a New Gliadin Capture Agent and Development of a Protein Microarray as a New Approach for Gliadin Detection

Cited by 4 publications

References 60 publications

Gluten Detection in Foods

Gluten Detection in Foods

Disposable electrochemical aptasensor for gluten determination in food

Label-Free Detection of Gliadin in Food by Quartz Crystal Microbalance-Based Immunosensor

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