Gliadin as a Measure of Gluten in Foods Containing Wheat, Rye, and Barley—Enzyme Immunoassay Method Based on a Specific Monoclonal Antibody to the Potentially Celiac Toxic Amino Acid Prolamin Sequences: Collaborative Study

Haas-Lauterbach, Sigrid; Cerne, V; Chirdo, Fernando Gabriel; Sadeleer, J de; Denery, Sandra; Feighery, Conleth; Hoertner, H; Höhne, Melanie; Hoinville, A; Iametti, Stefania; Immer, Ulrike; Janssen, Frederik; Koeppel, E; Yman, Ingrid Malmheden; Méndez, Enrique; Mothes, Thomas; Nissler, A; Raffler, G; Simón, Edurne; Thorell, Lars‐Håkan; Vela, Carmen; Vidts, A

doi:10.5740/jaoacint.cs2012_01

Cited by 23 publications

(16 citation statements)

References 3 publications

(3 reference statements)

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“…Several commercial enzyme-linked immunosorbent assay (ELISA) test kits suitable for the detection of gluten are available. At the time the gluten-free regulation was drafted, only the R5 monoclonal antibody-based and Morinaga Institutes of Biological Sciences (MIoBS) polyclonal antibody-based sandwich ELISAs (R5-sand and MIoBS-sand, respectively) had been validated by multiple laboratories at levels suitable for regulatory enforcement, extensively evaluated in research studies, and officially recognized by other governments or governmental agencies (1, 4 , 10,13,16,17,27). Since the 2013 gluten-free regulation, the G12 monoclonal antibody-based, A1-G12 monoclonal anti body-based, and Skerritt monoclonal antibody-based sandwich ELISAs (G 12-sand, A1-G 12-sand, and Skerrittsand, respectively) have been validated, extensively studied, and adapted to be suitable for detection of 20 ppm of gluten (2 , 18,19,21,22,25,28).…”

mentioning

confidence: 99%

Detection and Quantification of Gluten during the Brewing and Fermentation of Beer Using Antibody-Based Technologies

Panda

Zoerb

Cho

et al. 2015

Journal of Food Protection

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In 2013 the U.S. Food and Drug Administration (FDA) defined the term ''gluten-free'' and identified a gap in the analytical methodology for detection and quantification of gluten in foods subjected to fermentation and hydrolysis. To ascertain the ability of current enzyme-linked immunosorbent assays (ELISAs) to detect and quantify gluten in fermented and hydrolyzed products, sorghum beer was spiked in the initial phases of production with 0, 20, and 200 μg/ml wheat gluten, and samples were collected throughout the beer production process. The samples were analyzed using five sandwich ELISAs and two competitive ELISAs and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Western analysis employing four antibodies (MIoBS, R5, G12, and Skerritt). The sensitivity of the MIoBS ELISA (0.25 ppm) enabled the reliable detection of gluten throughout the manufacturing process, including fermentation, when the initial concentration of 20 μg/ml dropped to 2 μg/ml. The R5 antibody-based and G12 antibody-based sandwich ELISAs were unable to reliably detect gluten, initially at 20 μg/ml, after the onset of production. The Skerritt antibody-based sandwich ELISA overestimated the gluten concentration in all samples. The R5 antibody-based and G12 antibody-based competitive ELISAs were less sensitive than the sandwich ELISAs and did not provide accurate results for quantifying gluten concentration. The Western analyses were able to detect gluten at less than 5 μg/ml in the samples and confirmed the results of the ELISAs. Although further research is necessary before all problems associated with detection and quantification of hydrolyzed and fermented gluten are resolved, the analytical methods recommended by the FDA for regulatory samples can detect ≥ 20 μg/ml gluten that has undergone brewing and fermentation processes associated with the manufacture of beer.

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mentioning

confidence: 99%

Detection and Quantification of Gluten during the Brewing and Fermentation of Beer Using Antibody-Based Technologies

Panda

Zoerb

Cho

et al. 2015

Journal of Food Protection

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“…The limits of detection and quantification of the competitive R5 ELISA are 0.36 and 1.22 ng/ml of gliadins, respectively, being lower in liquid samples. Recently, a collaborative study has confirmed that the two R5 antibody-based ELISA test kits are able to detect gliadin at the lower level of the limit of detection with good reproducibility and repeatability (Immer and Haas-Lauterbach, 2012).…”

Section: Enzyme-linked Immunosorbent Assays (Elisas)mentioning

confidence: 99%

Cereals for developing gluten-free products and analytical tools for gluten detection

Rosell

Barro

Sousa

et al. 2014

Journal of Cereal Science

138

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Recently, gluten free foods have attracted much research interest motivated by the increasing market. Despite the motivation for developing gluten-free foods it is necessary to have a scientific basis for developing gluten-free foods and the tools for detecting the peptide sequence that could be immune-toxic to some persons. This review will be focused primarily on the cereal-based commodities available for developing gluten free blends, considering those naturally gluten free cereals besides the controversial oats, and the recent transgenic approaches for developing cereals free of immunotoxic gluten. Secondly, the biochemical tools for mimicking gluten network viscoelastic properties will be presented. Finally, special emphasis will be put in compiling the available techniques for gluten detection and quantitation.

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“…31,32 However, such quantitation may not give an accurate measure for other prolamins (secalin and hordein) and glutelins. Prolamins and glutelins differ in their proportions and molecular properties in wheat, rye and barley.…”

Section: ■ Introductionmentioning

confidence: 98%

Immunological Characterization of the Gluten Fractions and Their Hydrolysates from Wheat, Rye and Barley

Rallabhandi

Sharma

Pereira

et al. 2015

J. Agric. Food Chem.

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Gluten proteins in wheat, rye and barley cause celiac disease, an autoimmune disorder of the small intestine, which affects approximately 1% of the world population. Gluten is comprised of prolamin and glutelin. Since avoidance of dietary gluten is the only option for celiac patients, a sensitive gluten detection and quantitation method is warranted. Most regulatory agencies have set a threshold of 20 ppm gluten in foods labeled gluten-free, based on the currently available ELISA methods. However, these methods may exhibit differences in gluten quantitation from different gluten-containing grains. In this study, prolamin and glutelin fractions were isolated from wheat, rye, barley, oats and corn. Intact and pepsin-trypsin (PT)-digested prolamin and glutelin fractions were used to assess their immunoreactivity and gluten recovery by three sandwich and two competitive ELISA kits. The Western blots revealed varied affinity of ELISA antibodies to gluten-containing grain proteins and no reactivity to oat and corn proteins. ELISA results showed considerable variation in gluten recoveries from both intact and PT-digested gluten fractions among different kits. Prolamin fractions showed higher gluten recovery compared to their respective glutelin fractions. Among prolamins, barley exhibited higher recovery compared to wheat and rye with most of the ELISA kits used. Hydrolysis resulted in reduced gluten recovery of most gluten fractions. These results suggest that the suitability of ELISA for accurate gluten quantitation is dependent upon various factors, such as grain source, antibody specificity, gluten proteins and the level of their hydrolysis in foods.

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Gliadin as a Measure of Gluten in Foods Containing Wheat, Rye, and Barley—Enzyme Immunoassay Method Based on a Specific Monoclonal Antibody to the Potentially Celiac Toxic Amino Acid Prolamin Sequences: Collaborative Study

Cited by 23 publications

References 3 publications

Detection and Quantification of Gluten during the Brewing and Fermentation of Beer Using Antibody-Based Technologies

Detection and Quantification of Gluten during the Brewing and Fermentation of Beer Using Antibody-Based Technologies

Cereals for developing gluten-free products and analytical tools for gluten detection

Immunological Characterization of the Gluten Fractions and Their Hydrolysates from Wheat, Rye and Barley

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