Monoclonal Antibodies to High Molecular weight GluteninSubunits for Use in Measurement of Gluten in Foods

Background: Celiac disease (CD) is a heritable intolerance to gluten, the s t o r a g e protein found in wheat, rye, and barley. Commercially available assays u s e d t o c a l c u l at e g l u t e n c o nt e nt i n f o o d do not measure CD toxic high molecular weight glutenins (HMWG), but rely on the calculation that gliadin multiplied by two equals total gluten, which is invalid. Objective : We sought to generate monoclonal antibodies (mAbs), to known CD toxic HMWG subunits (HMW-GS), to develop improved assays to quantify CD toxic gluten in foods. Design: mAbs were generated using recombinant HMW-GS as antigens. They were termed CDC-3 and CDC-7 and were characterised by dot blot assay and ELISA. A suitable mAb was used in competitive ELISA to quantify HMW-GS content in foods intended to CD suffers. Results: Antibodies were raised against the important dough forming HMW-GS, 1Dy10 that cross-reacted with another important dough forming HMW-GS, 1Dx5. mAbs cross-reacted at high titre with HMW-GS extracted from fifteen varieties of European bread wheat. Competitive ELISA had a limit of detection of 625pg/ml, which is equivalent to 6.25x10 -3 ppm in a food extract. Conclusion: Our new antibodies react with a wide variety of wheat cultivars and can be used in sensitive ELISA assays for detection and quantification of CD toxic HMW-GS in foods intended to CD suffers. Summation of the food content of HMW-GS and gliadin permits improved quantitation of CD toxic gluten in nominally gluten-free foods.