QUANTITATIVE LC-MS DETECTION OF PEPTIDES IN CELIAC SPRUE

描述（由申请人提供）：乳糜泻是一种肠道遗传疾病，由摄入来自谷蛋白的免疫原性肽引起，谷蛋白是小麦、大麦和黑麦中存在的一类蛋白质，因此存在于许多市售食品中。症状很严重，唯一可行的治疗乳糜泻的方法是严格坚持无麸质饮食。本工作旨在为食品样品中免疫原性面筋肽的定性和定量检测的快速分析方法的发展提供支持。这种方法在未来对于无麸质食品的可靠认证是必要的，使乳糜泻患者能够以更低的时间和金钱要求保持他们的饮食。此外，这将有助于估计靶向破坏免疫原性肽的药物的有效性。这是特别有价值的，因为迄今为止还没有乳糜泻的动物模型。

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.