ELISA response and gliadin composition of different wheat cultivars grown in multiple harvest years

Hajas, Lívia; Scherf, Katharina Anne; Bugyi, Zsuzsanna; Török, Kitti; Schall, Eszter; Kohler, Peter O.; Tömösközi, Sándor

doi:10.1556/066.2016.0019

Cited by 12 publications

(11 citation statements)

References 27 publications

(29 reference statements)

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“…The wheat, rye, barley and oat flours were mixtures of four cultivars each to account for the genetic variability between different cultivars [ 38 ]. The cultivars were selected based on their production yields relative to the total production of winter wheat, rye, winter and summer barley, and oats in the year 2012 in Germany to include the most relevant cultivars (cumulative production share for wheat: 16%, rye: 53%, barley: 35%, oats: 41%) [ 39 ].…”

Section: Resultsmentioning

confidence: 99%

Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials

et al. 2017

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Gluten proteins from wheat, rye, barley and, in rare cases, oats, are responsible for triggering hypersensitivity reactions such as celiac disease, non-celiac gluten sensitivity and wheat allergy. Well-defined reference materials (RM) are essential for clinical studies, diagnostics, elucidation of disease mechanisms and food analyses to ensure the safety of gluten-free foods. Various RM are currently used, but a thorough characterization of the gluten source, content and composition is often missing. However, this characterization is essential due to the complexity and heterogeneity of gluten to avoid ambiguous results caused by differences in the RM used. A comprehensive strategy to isolate gluten protein fractions and gluten protein types (GPT) from wheat, rye, barley and oat flours was developed to obtain well-defined RM for clinical assays and gluten-free compliance testing. All isolated GPT (ω5-gliadins, ω1,2-gliadins, α-gliadins, γ-gliadins and high- and low-molecular-weight glutenin subunits from wheat, ω-secalins, γ-75k-secalins, γ-40k-secalins and high-molecular-weight secalins from rye, C-hordeins, γ-hordeins, B-hordeins and D-hordeins from barley and avenins from oats) were fully characterized using analytical reversed-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), N-terminal sequencing, electrospray-ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS) and untargeted LC-MS/MS of chymotryptic hydrolyzates of the single GPT. Taken together, the analytical methods confirmed that all GPT were reproducibly isolated in high purity from the flours and were suitable to be used as RM, e.g., for calibration of LC-MS/MS methods or enzyme-linked immunosorbent assays (ELISAs).

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Section: Resultsmentioning

confidence: 99%

Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials

et al. 2017

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“…To select marker peptides from wheat, a wheat flour mixture of four cultivars widely grown in Germany was used to include genetic variability between different cultivars [ 22 , 27 ]. The cultivars were selected based on their production yields relative to the total production of winter wheat, in the year 2012 in Germany to include the most relevant cultivars (cumulative production share for wheat: 16%) [ 28 ].…”

Section: Resultsmentioning

confidence: 99%

Targeted liquid chromatography tandem mass spectrometry to quantitate wheat gluten using well-defined reference proteins

2018

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Celiac disease (CD) is an inflammatory disorder of the upper small intestine caused by the ingestion of storage proteins (prolamins and glutelins) from wheat, barley, rye, and, in rare cases, oats. CD patients need to follow a gluten-free diet by consuming gluten-free products with gluten contents of less than 20 mg/kg. Currently, the recommended method for the quantitative determination of gluten is an enzyme-linked immunosorbent assay (ELISA) based on the R5 monoclonal antibody. Because the R5 ELISA mostly detects the prolamin fraction of gluten, a new independent method is required to detect prolamins as well as glutelins. This paper presents the development of a method to quantitate 16 wheat marker peptides derived from all wheat gluten protein types by liquid chromatography tandem mass spectrometry (LC-MS/MS) in the multiple reaction monitoring mode. The quantitation of each marker peptide in the chymotryptic digest of a defined amount of the respective reference wheat protein type resulted in peptide-specific yields. This enabled the conversion of peptide into protein type concentrations. Gluten contents were expressed as sum of all determined protein type concentrations. This new method was applied to quantitate gluten in wheat starches and compared to R5 ELISA and gel-permeation high-performance liquid chromatography with fluorescence detection (GP-HPLC-FLD), which resulted in a strong correlation between LC-MS/MS and the other two methods.

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“…To see whether harvest year or cultivar had a greater influence on 33-mer contents, four wheat cultivars (Mv Magvas, Mv Mazurka, Mv Verbunkos, and Yumai-34) grown at the same location in Hungary (Martonvásár) and harvested in three years (2011, 2012, and 2014) 35 were studied. The harvest year significantly influenced the 33-mer contents (p < 0.001), whereas the cultivars did not (p = 0.391).…”

Section: Resultsmentioning

confidence: 99%

Quantitation of the immunodominant 33-mer peptide from α-gliadin in wheat flours by liquid chromatography tandem mass spectrometry

Schalk

Lang

Wieser

et al. 2017

Sci Rep

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Coeliac disease (CD) is triggered by the ingestion of gluten proteins from wheat, rye, and barley. The 33-mer peptide from α2-gliadin has frequently been described as the most important CD-immunogenic sequence within gluten. However, from more than 890 published amino acid sequences of α-gliadins, only 19 sequences contain the 33-mer. In order to make a precise assessment of the importance of the 33-mer, it is necessary to elucidate which wheat species and cultivars contain the peptide and at which concentrations. This paper presents the development of a stable isotope dilution assay followed by liquid chromatography tandem mass spectrometry to quantitate the 33-mer in flours of 23 hexaploid modern and 15 old common (bread) wheat as well as two spelt cultivars. All flours contained the 33-mer peptide at levels ranging from 91–603 μg/g flour. In contrast, the 33-mer was absent (

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ELISA response and gliadin composition of different wheat cultivars grown in multiple harvest years

Cited by 12 publications

References 27 publications

Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials

Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials

Targeted liquid chromatography tandem mass spectrometry to quantitate wheat gluten using well-defined reference proteins

Quantitation of the immunodominant 33-mer peptide from α-gliadin in wheat flours by liquid chromatography tandem mass spectrometry

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