A modified extraction protocol enables detection and quantification of celiac disease-related gluten proteins from wheat

Broeck, H.C. van den; America, Twan; Smulders, M.J.M.; Bosch, Dirk; Hamer, R.J.; Gilissen, L.J.W.J.; Meer, I.M. van der

doi:10.1016/j.jchromb.2009.02.035

Cited by 65 publications

(27 citation statements)

References 44 publications

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“…For extraction of gluten proteins from wheat, we used the method as described by Van den Broeck et al (2009a), which combines three sequentially obtained extracts to extract nearly all gliadins and glutenins. Extraction of the residue left after the third extract with 25 mM Tris/HCl (pH 8.0) containing 2% SDS and with 25 mM Tris/HCl (pH 8.0) containing 2% SDS and 1% DTT showed that only some HMW-GS and probably some omega-gliadins/D-type LMW-GS were still present in the sample.…”

Section: Methodsmentioning

confidence: 99%

Presence of celiac disease epitopes in modern and old hexaploid wheat varieties: wheat breeding may have contributed to increased prevalence of celiac disease

Broeck¹,

Jong²,

Salentijn³

et al. 2010

Theor Appl Genet

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Gluten proteins from wheat can induce celiac disease (CD) in genetically susceptible individuals. Specific gluten peptides can be presented by antigen presenting cells to gluten-sensitive T-cell lymphocytes leading to CD. During the last decades, a significant increase has been observed in the prevalence of CD. This may partly be attributed to an increase in awareness and to improved diagnostic techniques, but increased wheat and gluten consumption is also considered a major cause. To analyze whether wheat breeding contributed to the increase of the prevalence of CD, we have compared the genetic diversity of gluten proteins for the presence of two CD epitopes (Glia-α9 and Glia-α20) in 36 modern European wheat varieties and in 50 landraces representing the wheat varieties grown up to around a century ago. Glia-α9 is a major (immunodominant) epitope that is recognized by the majority of CD patients. The minor Glia-α20 was included as a technical reference. Overall, the presence of the Glia-α9 epitope was higher in the modern varieties, whereas the presence of the Glia-α20 epitope was lower, as compared to the landraces. This suggests that modern wheat breeding practices may have led to an increased exposure to CD epitopes. On the other hand, some modern varieties and landraces have been identified that have relatively low contents of both epitopes. Such selected lines may serve as a start to breed wheat for the introduction of ‘low CD toxic’ as a new breeding trait. Large-scale culture and consumption of such varieties would considerably aid in decreasing the prevalence of CD.

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Section: Methodsmentioning

confidence: 99%

Presence of celiac disease epitopes in modern and old hexaploid wheat varieties: wheat breeding may have contributed to increased prevalence of celiac disease

Broeck¹,

Jong²,

Salentijn³

et al. 2010

Theor Appl Genet

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153

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“…The method of extraction of gliadin that is used herein resembles that used in the work of van den Broeck ( Van den Broeck et al, 2009). Initially, water-soluble proteins in a 1 g food sample were removed by shaking at 50 rpm in deionized water at room temperature for 1 h. After the water-soluble proteins had been extracted twice, globulins were extracted from the flour residue using 0.5 M NaCl solution.…”

Section: Pretreatment Of Samplementioning

confidence: 99%

Development of a barcode-style lateral flow immunoassay for the rapid semi-quantification of gliadin in foods

et al. 2016

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“…Classical methods for protein isolation from wheat grain are based on iso-propanol extraction (van den Broeck et al, 2009). We successfully implemented this method and have previously determined quantities of wheat grain proteins using gel-free proteomics approach (Uvackova et al, 2013a,b).…”

Section: Discussionmentioning

confidence: 99%

Using 7 cm immobilized pH gradient strips to determine levels of clinically relevant proteins in wheat grain extracts

et al. 2015

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The aim of the work was to test a relatively simple proteomics approach based on phenol extraction and two-dimensional gel electrophoresis (2-DE) with 7 cm immobilized pH gradient strips for the determination of clinically relevant proteins in wheat grain. Using this approach, 157 2-DE spots were quantified in biological triplicate, out of which 55 were identified by matrix-assisted laser desorption/ionization – time of flight tandem mass spectrometry. Clinically relevant proteins associated with celiac disease, wheat dependent exercise induced anaphylaxis, baker’s asthma, and food allergy, were detected in 24 2-DE spots. However, alcohol-soluble gliadins were not detected with this approach. The comparison with a recent quantitative study suggested that gel-based and gel-free proteomics approaches are complementary for the detection and quantification of clinically relevant proteins in wheat grain.

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A modified extraction protocol enables detection and quantification of celiac disease-related gluten proteins from wheat

Cited by 65 publications

References 44 publications

Presence of celiac disease epitopes in modern and old hexaploid wheat varieties: wheat breeding may have contributed to increased prevalence of celiac disease

Presence of celiac disease epitopes in modern and old hexaploid wheat varieties: wheat breeding may have contributed to increased prevalence of celiac disease

Development of a barcode-style lateral flow immunoassay for the rapid semi-quantification of gliadin in foods

Using 7 cm immobilized pH gradient strips to determine levels of clinically relevant proteins in wheat grain extracts

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