The production and characterisation of monoclonal antibodies to wheat gliadin peptides

Abstract

BALB/c mice maintained on a gluten-free diet were immunised with Frazer's Fraction III (FFIII, a peptic tryptic digest of wheat gluten that exacerbates coeliac disease) in order to overcome oral tolerance. A control group was maintained on normal diet. Serum antibody titres to FFIII were higher in the mice on a gluten-free diet ( P < 0.05). Three monoclonal antibodies to FFIII were produced from splenocytes obtained from mice maintained on a gluten-free diet. The antibodies were characterised by ELISA, immunodot assay and immunoblotting with prolamins from cereals toxic to coeliac patients (wheat, rye, barley, oats) and the non-toxic prolamins from maize and rice. The binding characteristics of the three antibodies to the cereal prolamins were different, implying that the antibodies recognise different cereal protein epitopes. Immunoblotting revealed FFIII to be comprised of antigenically dissimilar peptides.