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，并证明其用于经济诊断乳糜泻的实用性，乳糜泻是一种常见的自身免疫性消化疾病，与小麦、大麦和黑麦谷物中存在的麸质不耐受有关。此外，这项工作预计将导致更好地了解乳糜泻的原因。这项工作将导致识别大面板的肽配体，可以作为一个廉价的，但全面的诊断试剂面板腹腔疾病。将使用新的分子筛选方法鉴定这些肽。简而言之，该项目将涉及i）来自确诊的乳糜泻患者和健康对照组的血清组分的荧光标记，ii）针对混合血清抗体和耗尽HSA和IgG/伊加的总血清蛋白的细菌展示肽文库的基于特异性的筛选，iii）鉴定文库选择的克隆用于诊断阵列构建，和iv）训练阵列以使相对于对照组的诊断特异性和灵敏度最大化。然后，将使用出现CD症状的未确诊患者组验证诊断阵列。公共卫生相关性：现有的用于询问存在于循环库中的不同抗体特异性的方法已经产生了对自身免疫性疾病和癌症的新的见解和诊断。所提出的方法可以扩展这些能力，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。这项工作将建立一个新的金标准方法，用于分析血清抗体特异性，以及提供试剂来诊断常见的自身免疫性疾病（乳糜泻），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.