Quantitative Serum Antibody Specificity Screening in Celiac Disease

描述（由申请人提供）：这项工作的主要目的是开发一种蛋白质组学分析方法，并证明其在乳糜泻（一种常见的自身免疫性消化疾病，与小麦、大麦和黑麦谷物中的麸质不耐受有关）经济诊断中的应用。此外，这项工作有望使人们更好地了解乳糜泻的病因。这项工作将导致鉴定出大量的肽配体，这些肽配体可以作为乳糜泻廉价但全面的诊断试剂。这些肽将使用一种新的分子筛选方法进行鉴定。简而言之，该项目将包括i)对确诊乳糜泻患者和健康对照组的血清成分进行荧光标记，ii)基于特异性的细菌展示肽文库筛选，以对抗汇集的血清抗体和HSA和IgG/IgA的血清总蛋白，iii)鉴定文库选择的克隆用于诊断阵列的构建，以及iv)对阵列进行培训，以最大限度地提高诊断特异性和相对于对照组的敏感性。然后，诊断阵列将在出现CD症状的未确诊患者组中进行验证。公共卫生相关性：用于询问循环库中存在的各种抗体特异性的现有方法已经为自身免疫性疾病和癌症提供了新的见解和诊断。提出的方法可以扩展这些功能，提供更多关于疾病病理学的信息，并允许生成改进的生化试剂，以创建更有效但更便宜的诊断测试。这项工作将为血清抗体特异性分析建立一个新的金标准方法，并为诊断一种常见的自身免疫性疾病（乳糜泻）提供试剂，并更好地了解其原因。

DESCRIPTION (provided by applicant): The primary objective of this work is to develop a proteomic analysis method and demonstrate its utility for economical diagnosis of Celiac Disease, a common autoimmune digestive disorder associated with intolerance to gluten present in wheat, barley, and rye grains. In addition, this work is expected to lead to a better understanding of the cause of Celiac Disease. This work will result in the identification large panel of peptide ligands that can serve as an inexpensive but comprehensive diagnostic reagent panel for Celiac Disease. These peptides will be identified using a new molecular screening methodology. Briefly, this project will involve i) fluorescent- labeling of serum components from patients with confirmed Celiac Disease and healthy control groups, ii) specificity-based screening of bacterial display peptide libraries against pooled serum antibody and total serum protein depleted of HSA and IgG/IgA, iii) identification of library selected clones for diagnostic array construction, and iv) training of the array to maximize diagnostic specificity and sensitivity relative to the control group. The diagnostic array will then be validated with an undiagnosed patient group presenting the symptoms of CD. PUBLIC HEALTH RELEVANCE: Existing methodologies for interrogating the diverse antibody specificities present in the circulating repertoire have yielded new insights and diagnostics for autoimmune diseases and cancer. The proposed approach could expand these capabilities providing greater information about disease pathology, and allowing for the generation of improved biochemical reagents to create more effective yet inexpensive diagnostic tests. This work will establish a new gold standard methodology for analysis of serum antibody specificities, as well as provide reagents to diagnose a common autoimmune disorder (Celiac Disease), and better understand its cause.