QUANTITATIVE LC-MS DETECTION OF PEPTIDES IN CELIAC SPRUE

DESCRIPTION (provided by applicant): Celiac Sprue is an inherited disease of the intestine caused by ingestion of immunogenic peptides derived from gluten, a class of proteins present in wheat, barley and rye and therefore in many commercially available foods. Symptoms are severe, the only feasible treatment for Celiac Sprue being a strict adherence to a gluten free diet. This work seeks support for the development of a rapid analytical method for the qualitative and quantitative detection of immunogenic gluten peptides from food samples. Such a method will be necessary in the future for a reliable certification of gluten free foods, enabling Celiac patients to keep their diet with lower time and money requirements. Furthermore, it will be useful to estimate the affectivity of drugs targeted to destroy immunogenic peptides. This is particularly valuable because no animal model for Celiac Sprue exists to date. The method envisioned will be based on Liquid Chromatography coupled Mass Spectrometry (LC-MS), having the advantage of high throughput allowing low operating costs and easy transferability. Research will first focus on identification of known immunogenic fragments from gluten by LC/electrospray- MS/MS. The peptides will be between 10 and 40 amino acids in length, and ionization as well as fragmentation conditions have to be carefully chosen to achieve maximum reliability of sequence assignments. The marker gluten peptides will be chosen based on relative abundance and unique to immunogenic celiac peptides in grains such as wheat, barley and rye while absent in non immunogenic grains such as rice and corn In a parallel effort, a small number of representative peptides will be chemically synthesized and used as internal standards for quantification of the gluten marker peptides. In a potential Phase II project, the method will be optimized for robustness and general applicability, and broadened in the number of peptides recognized. The efforts will pave the way for eventual commercial application as a service to both the Celiac community and the food industry, and provide a simple and economic way to safe and affordable food for Celiac patients.