Monoclonal antibodies to high molecula weight glutenin subunits for use in measurement of gluten in foods

Identificador:  imarina:5129533

Handle: https://hdl.handle.net/20.500.11797/imarina5129533

Autors:  H Julia Ellis; M Carmen Bermudo Redondo; Tanja Suligoj; Nika Japelij; Nuria Gonzalez-Cinca; Beatriz Côrte-Real; Paul J Ciclitira

Resum:
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 used to calculate gluten content in food 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.