Quantitative Serum Antibody Specificity Screening in Celiac Disease

描述(申请人提供)：这项工作的主要目标是开发一种蛋白质组学分析方法，并证明其对乳糜泻的经济诊断的有效性。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面筋不耐受有关。此外，这项工作有望使人们更好地了解腹腔疾病的病因。这项工作将导致鉴定大量的多肽配体，这些多肽配体可以作为一种廉价但全面的腹腔疾病诊断试剂板。这些多肽将使用一种新的分子筛选方法进行鉴定。简而言之，这个项目将包括：i)来自确诊的乳糜泻患者和健康对照组的血清成分的荧光标记，ii)针对混合血清抗体和总血清蛋白的细菌展示肽库的特异性筛选，iii)鉴定用于诊断阵列构建的文库选择克隆，以及iv)训练阵列以最大限度地提高相对于对照组的诊断特异性和灵敏度。诊断阵列随后将与出现CD症状的未诊断患者组进行验证。与公共卫生相关：现有的方法用于询问循环曲目中存在的各种抗体特异性，为自身免疫性疾病和癌症提供了新的见解和诊断方法。拟议的方法可以扩展这些能力，提供更多关于疾病病理的信息，并允许产生改进的生化试剂，以创建更有效但更便宜的诊断测试。这项工作将建立一种新的分析血清抗体特异性的金标准方法，并为诊断一种常见的自身免疫性疾病(腹腔疾病)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.