Characterizing and optimizing IgE and IgG4 microarray peptide assays for Ara h 2

表征和优化 Ara h 2 的 IgE 和 IgG4 微阵列肽测定

• 批准号: 10289505
• 负责人: STEPHEN C DRESKIN
• 金额: $ 7.78万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-07 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10289505
• 关键词: 16 year old; Acute; Address; Affect; Albumins; Allergens; Allergic; Allergic Reaction; Allergy to peanuts; American; Anaphylaxis; Angioneurotic Edema; Anxiety; Binding; Biological Assay; Bronchial Spasm; Caregivers; Cessation of life; Child; Clinical; Clinical Trials; Data; Diagnostic; Enzyme-Linked Immunosorbent Assay; Epitopes; Face; Food; Food Hypersensitivity; Health; Health Care Costs; IgE; IgG4; Immunoassay; Immunoglobulins; Infant; Knowledge; Measurement; Measures; Mediating; Northern Europe; Outcome Study; Patients; Pattern; Peptides; Persons; Population; Predictive Value; Prevalence; Process; Proteins; Public Health; Quality of life; Reaction; Reproducibility; Risk; Sampling; Severities; Swelling; Testing; United States; Urticaria; base; clinically relevant; improved; mast cell; negative affect; oral immunotherapy; screening; tool

Project Summary: Peanut allergy is recognized as one of the most severe food allergies due to its prevalence, persistence, and the potential severity. Peanut allergy reactions vary from mild to severe, ranging from acute hives, severe angioedema, swelling of the face, bronchospasm, anaphylaxis, to even death. The US annual healthcare cost of managing peanut allergy is estimated to be several billion dollars. Thus, there is an urgent need to develop improved diagnostics. Among the 17 known PN allergens, the highly homologous 2S albumins, Ara h 2 and Ara h 6, are the most potent for eliciting IgE-mediated mast cell activation and measurement of specific IgE to either Ara h 2 or Ara h 6 correlates best with clinically relevant PN allergy. Peptide microarrays have been in use for a decade and are particularly useful for screening peptides and qualitative analysis of immunoglobulin binding. We hypothesize that these peptide microarrays have the potential to be more quantitative and become important diagnostics but there are important gaps in knowledge in regards to the size of the peptides assayed, how samples are handled, sensitivity, reproducibility and the dynamic range of the assays. We will use samples from a well characterized clinical trial of oral immunotherapy to test the following hypotheses: 1) normalizing sera based on PN-sIgE (or IgG4) or Ara h 2-sIgE (or IgG4) prior to assay will give more accurate measure of quantitative binding to peptides that may be seen if sera are assayed at a fixed dilution and 2) microarray assays are more sensitive than either Bt-ELISA or ImmunoCap®, but have a more restricted dynamic range. The outcome of these studies will be an enhanced understanding of the use of microarrays so that this important tool can be applied to a variety of allergic conditions.

项目摘要： 由于其患病率，花生过敏被认为是最严重的食物过敏之一， 持久性和潜在的严重性。花生过敏反应从轻度到重度不等 从急性蜂箱，严重的血管性水肿，脸部肿胀，支气管痉挛，过敏反应到甚至 死亡。美国管理花生过敏的年度医疗保健费用估计为几个 十亿美元。这是迫切需要开发改进的诊断。在17中 已知的PN过敏原，高度同源的2S专辑ARA H 2和ARA H 6是最多的 引发IgE介导的肥大细胞激活和特异性IgE的测量的潜力 ARA H 2或ARA H 6与临床上相关的PN过敏最为相关。肽微阵列具有 使用了十年，对于筛查肽和定性分析特别有用 免疫球蛋白结合。我们假设这些Pepperide微阵列有潜力 更定量并成为重要的诊断，但存在重要差距 关于分配的宠物大小的知识，如何处理样本，敏感性， 可重复性和测定的动态范围。我们将使用井中的样品 表征口服免疫疗法的临床试验以检验以下假设：1）正常化 基于PN-SIGE（或IgG4）或ARA H 2-SIGE（或IgG4）之前的血清将提供更准确的 定量结合与辣椒的定量结合度量，如果在固定稀释下分配血清 2）微阵列测定比BT-Elisa或ImmunoCap®更敏感，但具有 动态范围更大。这些研究的结果将是一种增强的理解 使用微阵列，以便将此重要的工具应用于多种过敏性 状况。