Gliadin analysis by reversed-phase high performance liquid chromatography

Gojković-Cvjetković, S Vesna; Grujić, D Radoslav; Marjanović-Balaban, R Željka; Stanojevic, P Ljiljana; Stanojević, S Jelena; Cakic, D Milorad

doi:10.5937/savteh1902030g

Cited by 7 publications

(7 citation statements)

References 16 publications

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“…The applied RP-HPLC method was carried out according to Gojković-Cvjetković et al [ 23 ] with the following modifications. The contents and characterization of the isolated avenins were assessed in samples (10 µL) through reverse-phase high-performance liquid chromatography (RP-HPLC) using a Waters 2965 apparatus with both a UV detector (210 nm) and 300 SB-C8 Zorbax Poroshell ™ column (75 mm × 2.1 mm, 5 µm particles) linked to a Zorba 300SB-C8 cartridge guard column (Rockland Technologies, Inc., Newport, DE, USA).…”

Section: Methodsmentioning

confidence: 99%

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Dvořāček

Kotrbová-Kozak

Kozová-Doležalová

et al. 2022

Foods

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Current clinical studies confirm that the consumption of oats for people suffering from celiac disease is safe. Some studies have confirmed different levels of immunoreactive gluten epitopes of oats in different cultivars, while others explain these differences due to contamination with gluten-rich species or as random cross-reactivity ELISA of homologous oat epitopes with anti-wheat gliadin antibodies. The aim of our two-year study was therefore to map cross-reactive oat epitopes in a set of 132 oat cultivars using a G12-based ELISA kit. The results were focused on the varietal and annual level of cross-reactivity (interference) of avenin epitopes with the G12 antibody on the identification of potential cultivars with significantly different interferences and assessing the degree of risk of possible false-contamination with external gluten. Although repeated evaluations confirmed high year-to-year variability (RSD ≥ 30%) in approximately 2/3 of the cultivars, the content of interfering avenin epitopes with G12 did not exceed the considered safe limit (20 mg·kg−1) for celiacs. At the same time, not only annual but, above all, significant cultivar dependences in the interference of avenins to the G12 antibody were demonstrated. Genetic dependence was further confirmed in connection with the proven avenin polymorphism as well as immunoblotting with the identification of interfering peptides with the G12 antibody in the 25 and 30 kDa regions. It was the occurrence of two bands around 30 kDa that predominantly occurred in oat cultivars with a relatively higher content of cross-reactive avenins (12–16 mg·kg−1). Due to the fact that the contents of interfering avenins ranged in several cultivars even over 16 mg·kg−1, the choice of a suitable oat cultivar may be crucial for gluten-free food producers, as it reduces the risk of a possible false-response of the commercial ELISA kits when checking the real-gluten contamination.

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Section: Methodsmentioning

confidence: 99%

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Dvořāček

Kotrbová-Kozak

Kozová-Doležalová

et al. 2022

Foods

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“…The gliadin and glutenin extracts, extracted from the above‐mentioned wheat flour, were obtained according to the modified method of Wieser et al (1988) and Gojković Cvjetković et al (2019). The extracts were taken from 100 mg wheat flour samples.…”

Section: Methodsmentioning

confidence: 99%

“…These are deionized water obtained under laboratory conditions (on the apparatus Siemens water Technologies W3T199551, Siemens Ultra Clear, Germany, at a conductivity of 0.055 mS/cm and at a temperature of 20 ℃, eluent A) and 0.1% trifluoroacetic acid (TFA, Lach‐Ner, Czech Republic, high purity, 99.50%), in acetonitrile (ACN, Lach‐Ner, Czech Republic, high purity, 99.95%), (eluent B). Separation was performed according to the modified method of Gojković Cvjetković et al (2019). The results were processed using the Agilent OpenLab CDS ChemStation software.…”

Section: Methodsmentioning

confidence: 99%

Investigation of the effect of cold atmospheric plasma on gliadins and glutenins extracted from wheat flour samples

Cvjetković

Marjanović-Balabań

Vujadinović

et al. 2021

Food Processing Preservation

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Gluten proteins a type of storage protein in wheat (Triticum aestivum L.), indispensable for the development of the gluten network in dough, contain two fractions: gliadins and glutenins (Shewry, 2009(Shewry, , 2019. Gliadins are monomeric molecules, and glutenins form an extended network, synergistically forming the gluten network (Balakireva & Zamyatnin, 2016;Shewry & Lookhart, 2003).These proteins have great technological importance in many food systems (Delcour et al., 2012;Rustgi et al., 2019). They are responsible for the rheological and viscoelastic properties of wheat dough (Mioduszewski & Cieplak, 2021;Shewry, 2009). Gliadin fraction affects the extensibility, while glutenin fraction affects the strength and elasticity of the wheat dough (Mioduszewski & Cieplak, 2021).A great number of food allergens are proteins whose molecular weight ranges from 10 to 70 kDa (Besler et al., 2001;Monaci et al., 2020). Different biological, chemical, and physical approaches have been applied for the degradation of these proteins (Valerii et al.,

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“…The applied RP-HPLC method were carried out according to Gojković-Cvjetković et al [17] with following modifications. Contents and characterization of isolated avenins were assessed in samples (10 µl) through reverse-phase high performance liquid chromatography (RP-HPLC) using a Waters 2965 apparatus with UV detector (210 nm) and 300 SB-C8 Zorbax Poroshell ™ column (75×2.1 mm, 5 µm particles) linked to a Zorba 300SB-C8 cartridge guard column (RocklandTechnologies, Inc.Newport, DE).…”

Section: Avenin Quantification By Rp-hplcmentioning

confidence: 99%

Identification of Perspective Oat Cultivars with a Minimum Content of Gluten Homologous Peptides

Dvořāček¹,

Kotrbová-Kozak²,

Kozová-Doležalová³

et al. 2020

Preprint

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There is still an ambiguous opinion on oat regarding its safety for people with celiac disease (CD). Some studies have confirmed different content of oat immunoreactive epitopes in different cultivars while others explain the differences in consequence of cross-contamination with gluten-rich species or as ELISA cross-reactivity of oat homological epitopes with antibodies against wheat gliadin. Our study was based on a two-year mapping of oat immunoreactive epitopes in a set of 132 oat cultivars using the G12-based ELISA kit. Although repeated evaluation confirmed high interannual variability (RSD ≥ 30%) in approximately 2/3 of the cultivars, the permitted gluten content (20 mg kg-1) has not been exceeded except for contaminated cultivar Sirene. The polymorphism of purified avenins determined by SDS-PAGE revealed the occurrence of 2 bands around 30 kDa in oat cultivars with relatively high gluten content (12-16 mg kg-1) except for the cultivar Leo while this pattern occurred only in 50% oat cultivars with low gluten content. Quantification of gluten epitopes on purified avenin unit further revealed the three materials (Mojacar, Maris Oberon, SG-K 16370) with the lowest gluten content and presence of one band at 30 kDa. The band pattern at 30 kDa thus represents a promising breeding marker.

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Gliadin analysis by reversed-phase high performance liquid chromatography

Cited by 7 publications

References 16 publications

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Investigation of the effect of cold atmospheric plasma on gliadins and glutenins extracted from wheat flour samples

Identification of Perspective Oat Cultivars with a Minimum Content of Gluten Homologous Peptides

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