QUANTITATIVE LC-MS DETECTION OF PEPTIDES IN CELIAC SPRUE

乳糜泻中肽的定量 LC-MS 检测

• 批准号: 6832089
• 负责人: ROBERT D VOYKSNER
• 金额: $ 10万
• 依托单位: LCMS LIMITED
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-27 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6832089
• 关键词: celiac disease; corn; electrospray ionization mass spectrometry; enzyme linked immunosorbent assay; gas chromatography mass spectrometry; high throughput technology; immunotoxicity; liquid chromatography mass spectrometry; peptides; plant proteins; protein glutamine gamma glutamyltransferase; proteolysis; rice; technology /technique development; tissue /cell culture; wheat

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.

描述（由申请人提供）：乳糜泻是一种肠道遗传性疾病，由摄入来源于麸质的免疫原性肽引起，麸质是小麦、大麦和黑麦中存在的一类蛋白质，因此也存在于许多市售食品中。症状是严重的，唯一可行的治疗乳糜泻是严格遵守无麸质饮食。这项工作寻求支持的发展，定性和定量检测食品样品中的免疫原性谷蛋白肽的快速分析方法。这种方法将是必要的，在未来的一个可靠的认证无麸质食品，使乳糜泻患者保持他们的饮食与较低的时间和金钱的要求。此外，这将有助于评估靶向破坏免疫原性肽的药物的有效性。这是特别有价值的，因为没有动物模型的腹腔Sprue存在的日期。 所设想的方法将基于液相色谱-质谱联用（LC-MS），具有高通量的优点，允许低操作成本和易于转移。研究将首先集中于通过LC/电喷雾- MS/MS鉴定来自谷蛋白的已知免疫原性片段。肽的长度将在10至40个氨基酸之间，并且必须仔细选择电离以及片段化条件以实现序列分配的最大可靠性。标记物谷蛋白肽将基于相对丰度来选择，并且对于谷物如小麦、大麦和黑麦中的免疫原性腹腔肽是独特的，而在非免疫原性谷物如稻和玉米中不存在。在平行的努力中，将化学合成少量代表性肽并用作定量谷蛋白标记物肽的内标。 在一个潜在的II期项目中，该方法将针对耐用性和普遍适用性进行优化，并扩大识别肽的数量。这些努力将为最终的商业应用铺平道路，为乳糜泻社区和食品行业提供服务，并为乳糜泻患者提供安全和负担得起的食品提供简单经济的方式。