Molecular and Immunological Characterization of Gluten Proteins Isolated from Oat Cultivars That Differ in Toxicity for Celiac Disease

Real, Ana; Comino, Isabel; Lorenzo, Laura de; Merchán, Francisco; Gil-Humanes, Javier; Giménez, María J.; López-Casado, Miguel Ángel; Torres, Ma Isabel; Cebolla, Ángel; Sousa, Carolina; Barro, Francisco; Pistón, Fernando

doi:10.1371/journal.pone.0048365

Cited by 76 publications

(95 citation statements)

References 42 publications

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“…Compared to other cereals, grains belonging to subtribe Triticeae (wheat, barley, and rye) contain significantly higher levels of glutamine and proline than others, being these amino acids the principal responsible for triggering the immune response in celiac disease [25]. A direct correlation between the immunogenicity of the different oat varieties and the presence of specific peptides with differential reactivities has been proposed as the origin of the wide range of variation of potential immunotoxicity of oat cultivars [40].…”

Section: Celiac Disease (Cd)mentioning

confidence: 99%

Determination of Gluten Peptides Associated with Celiac Disease by Mass Spectrometry

Alves¹,

D’Almeida²,

Ferreira³

2017

Celiac Disease and Non-Celiac Gluten Sensitivity

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Gluten is a big protein network composed of monomeric fraction (prolamins) and polymeric fraction (glutelins), occurring in many cereal-based products, especially in those containing wheat. Gluten peptides can trigger food allergies and intolerances, including inflammatory reactions as the celiac disease, an autoimmune disorder of the small intestine characterized by mucosal degeneration and villous atrophy. The treatment is the permanent exclusion of gluten from diet. However, gluten analysis is a very difficult task, due to the high complexity of polypeptides and the lack of consensus on the most appropriate analytical method. Proteomics approaches, combining liquid chromatography and mass spectrometry in tandem (LC-MS/MS), have been pointed as the most promising nonimmunological techniques for gluten detection. LC-MS analyses associated with bioinformatics and specific-prolamin database can solve methodological limitations since it is based on the accurate molecular mass of peptide biomarkers. One of the major contributions of proteomics has been the identification of epitopes of gluten peptides responsible for wheat-related diseases. Recent works have defined grain-specific gluten peptides and also the lowest concentration at which peptides could be confidently detected. Proteomic application for gluten quantification should support not only regulatory limits in processed foods, but also the safety of consumers about food labeled as gluten-free.

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Section: Celiac Disease (Cd)mentioning

confidence: 99%

Determination of Gluten Peptides Associated with Celiac Disease by Mass Spectrometry

Alves¹,

D’Almeida²,

Ferreira³

2017

Celiac Disease and Non-Celiac Gluten Sensitivity

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show abstract

“…However, the toxicity of oat for coeliac patients is still controversial [24]( [25] and citation therein), so oat prolamins also have to be evaluated towards the quantification of the toxicity in food products. For this reason it is necessary to evaluate the response of the competitive assays using Gli 1 and Gli 4 aptamers against prolamins from other toxic cereals and the selectivity against not-CDtriggering cereals and grains.…”

Section: Cross-reactivity Against Other Cereals and Grainsmentioning

confidence: 99%

Sensitive gluten determination in gluten-free foods by an electrochemical aptamer-based assay

Amaya-González

de‐los‐Santos‐Álvarez

Miranda‐Ordieres

et al. 2015

Anal Bioanal Chem

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Enzyme-immune assays are currently the methods of choice for gluten control in foods labelled as "gluten free", providing a mechanism for assessing food safety for consumption by coeliac and other allergic patients. However, their limitations, many of them associated to the reactivity of the different antibodies used and their degree of specificity, have prevented the establishment of a standardized method of analysis. We explore new methods for quantitatively determine gluten content in foods based on the use of two recently described aptamers, raised against a 33-mer peptide recognised as the immunodominant fragment from α2-gliadin. The assays use the target peptide immobilized onto streptavidin-coated magnetic beads in combination with a limited amount of biotin-aptamer in a competitive format, followed by streptavidin-peroxidase labelling of the aptamer that remains bound to the magnetic beads. The enzyme activity onto the beads, measured by chronoamperometry in disposable screen-printed electrodes, is inversely related to the target concentration in the test solution. We find that while the assay using the aptamer with the highest affinity towards the target (Gli 4) achieves low detection limits (~0.5 ppm) and excellent analytical performance when challenged in samples containing the intact protein, gliadin, it fails in detecting the peptide in solution. This problem is circumvented by employing another aptamer (Gli 1), the most abundant one in the SELEX pool, as a receptor. The proposed assays allow the convenient detection of the allergen in different kind of food samples, including heat-treated and hydrolysed ones. The obtained results correlate with those of commercially available antibody-based assays, providing an alternative for ensuring the safety and quality of nominally gluten-free foods.

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“…These fractions were subjected to pepsin, trypsin and chymotrypsin sequential digestion, as previously described by Real et al [3].…”

Section: Methodsmentioning

confidence: 99%

Identification and In Vitro Reactivity of Celiac Immunoactive Peptides in an Apparent Gluten-Free Beer

et al. 2014

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Gluten content from barley, rye, wheat and in certain oat varieties, must be avoid in individuals with celiac disease. In most of the Western countries, the level of gluten content in food to be considered as gluten-free products is below 20 parts per million measured by ELISA based on specific anti-gluten peptide antibody. However, in beverages or food suffering complex hydrolytic processes as beers, the relative proportion of reactive peptides for celiac patients and the analytical techniques may differ, because of the diversity of the resulting peptide populations after fermentations. A beer below 20 parts per million of gluten but yet detectable levels of gluten peptides by anti-gliadin 33-mer antibodies (G12 and A1) was analyzed. We identified and characterized the relevant peptides for either antibody recognition or immunoactivity in celiac patients. The beer was fractionated by HPLC. The relative reactivity of the different HPLC fractions to the G12/A1 antibodies correlated to the reactivity of peripheral blood mononuclear cells isolated from 14 celiac individuals. Peptides from representative fractions classified according to the relative reactivity to G12/A1 antibodies were identified by mass spectrometry. The beer peptides containing sequences with similarity to those of previously described G12 and A1 epitopes were synthesized and confirmed significant reactivity for the antibodies. The most reactive peptides for G12/A1 also confirmed the highest immunogenicity by peripheral blood mononuclear cell activation and interferon γ production from celiac patients. We concluded that preparative HPLC combined with anti-gliadin 33-mer G12/A1 antibodies were very sensitive and specific methods to analyze the relevant immunogenic peptides in hydrolyzed gluten.

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Molecular and Immunological Characterization of Gluten Proteins Isolated from Oat Cultivars That Differ in Toxicity for Celiac Disease

Cited by 76 publications

References 42 publications

Determination of Gluten Peptides Associated with Celiac Disease by Mass Spectrometry

Determination of Gluten Peptides Associated with Celiac Disease by Mass Spectrometry

Sensitive gluten determination in gluten-free foods by an electrochemical aptamer-based assay

Identification and In Vitro Reactivity of Celiac Immunoactive Peptides in an Apparent Gluten-Free Beer

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