Sensitive detection of cereal fractions that are toxic to celiac disease patients by using monoclonal antibodies to a main immunogenic wheat peptide

Abstract

Both ELISAs designed for use with the toxic gliadin 33-mer peptide suggested a high correlation between the presence of the peptide and the amount of cereal that was toxic to celiac disease patients. The sensitivity was significantly higher than that of equivalent methods recognizing other gluten epitopes.

Improved extraction of prolamins for gluten detection in processed foods

“…The safety of gluten-free products for people with gluten intolerance is controlled by gluten enzyme-linked immunosorbent assays (ELISA) (e.g. Skerritt and Hill 1990, Valdés et al 2003, Morón et al 2008). These gluten methods are able to detect very low levels of prolamin proteins, and new limits for acceptable gluten contents in gluten-free products have been set by the European Union (Commission regulation (EC) No 41/2009).…”

Detection of Allergen Markers in Food: Analytical Methods

“…Hence the ELISA methods used for gluten detection in foods utilize antibodies that bind to common gluten epitopes found in wheat, rye, and barley. Some of the well-characterized monoclonal antibodies used in commercial ELISA kits include Skerritt or 401/21 [97], R5 [98], and G12 [99]. The variable reactivity of these anti-gluten Th anti bodies towards gluten from diff erent grain sources of wheat, rye, and ff ff barley may result in under-or overestimation of gluten in foods [100,101].…”

Sensitive detection of cereal fractions that are toxic to celiac disease patients by using monoclonal antibodies to a main immunogenic gluten peptide

“…Two monoclonal antibodies (moAbs), A1 and G12, were raised against the immunodominant peptide 33-mer (LQLQPFPQPQLPYPQPQLPYPQPQLPYP QPQPF, residues 57 to 89; Morón et al, 2008a).…”

“…Thus the production of moAbs against this toxic gluten peptide could be of great importance in both research and diagnosis. We obtained moAbs against the 33-mer peptide (A1 and G12 moAbs) (Morón et al, 2008a). To test the relative sensitivity of each moAb for the 33-mer peptide, we immobilized different concentrations of the C-LYTAG-33-mer polypeptide, and detected with A1 and G12 moAbs in an indirect ELISA.…”

“…We investigated whether the G12/A1 moAbs were able to detect the presence of gliadin 33-mer-related epitopes in prolamines from various cereals (Morón et al, 2008a;2008b). To obtain quantitative data on the capacity of these antibodies to detect coeliac-toxic prolamines, we performed an indirect ELISA with samples of wheat, barley, rye, oats, rice, and maize (Figure 2).…”