Quantitative Mass Spectrometric Immunoassays for Population Proteomics

群体蛋白质组学的定量质谱免疫分析

• 批准号: 7744570
• 负责人: DOBRIN NEDELKOV
• 金额: $ 19.73万
• 依托单位: INTRINSIC BIOPROBES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2010-09-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7744570
• 关键词: Animals; Antibodies; Benchmarking; Biological Assay; Characteristics; Cohort Studies; Collection; Data; Detection; Development; Discipline; Disease; Disease Progression; Enzyme-Linked Immunosorbent Assay; Exhibits; Frequencies; General Population; Goals; Grant; Hand; Human; Immunoassay; Individual; Investigation; Modification; Monitor; Non-Human Protein; Pathologic Processes; Pattern; Phase; Plasma; Plasma Proteins; Population; Post-Translational Protein Processing; Prealbumin; Protein Isoforms; Proteins; Proteomics; Reference Standards; Retinol Binding Proteins; Role; Sampling; Sensitivity and Specificity; Specificity; Structural Protein; Time; Transferrin; Validation; Variant; assay development; base; beta-2 Microglobulin; cohort; cross reactivity; disease diagnosis; insight; post gamma-globulins; public health relevance; response

DESCRIPTION (provided by applicant): The main objective of this Phase I project is to develop quantitative Mass Spectrometric Immunoassays for beta-2-microglobulin, cystatin C, transferrin, transthyretin, and retinol binding protein. These five proteins were recently assayed via qualitative mass spectrometric immunoassays in 1,000 healthy cohorts, detecting (without providing concentration information) numerous protein variant. Because sandwich-based immunoassays such as ELISAs cannot specifically quantify the subtle protein modifications, in this project we propose to develop quantitative mass spectrometric immunoassays for these proteins and their isoforms. An internal reference standard will be carefully chosen, and a step-by-step standard curve quantitative mass spectrometric immunoassay development will be undertaken. The precision (intra-assay and inter-assay), linearity, sensitivity, and specificity of the quantitative mass spectrometric immunoassays will be determined. As a final validation step, the quantitative MSIA assays will be evaluated and benchmarked against existing ELISA assays. The quantitative mass spectrometric immunoassays proposed here will offer unique and straightforward means of quantifying protein variants, and when applied to disease cohorts, can provide valuable insight into the role of these variants in the onset of the disease, progression, and response to therapy. PUBLIC HEALTH RELEVANCE: The development of quantitative Mass Spectrometric Immunoassays for beta-2- microglobulin, cystatin C, transferrin, transthyretin, and retinol binding protein will enable routine quantitative monitoring of these proteins and the changes in their modifications patterns that might be relevant to disease. With the new assays in hand, we will be better equipped to study the effects of these proteins' modifications in pathological processes and evaluate their potential as new indicators of the onset and progression of diseases, and response to therapy.

描述（由申请方提供）：本I期项目的主要目标是开发β 2-微球蛋白、胱抑素C、转铁蛋白、甲状腺素运载蛋白和视黄醇结合蛋白的定量质谱免疫测定。这五种蛋白质最近通过定性质谱免疫测定法在1,000个健康队列中进行了测定，检测（不提供浓度信息）许多蛋白质变体。由于基于ELISA的免疫测定法，如ELISA不能具体量化的微妙的蛋白质修饰，在这个项目中，我们建议开发定量质谱免疫测定这些蛋白质及其亚型。将仔细选择内部参比标准品，并逐步进行标准曲线定量质谱免疫测定开发。将测定定量质谱免疫测定的精密度（试验内和试验间）、线性、灵敏度和特异性。作为最终验证步骤，将根据现有ELISA检测法对定量MSIA检测法进行评价和基准化。本文提出的定量质谱免疫测定将提供独特而直接的蛋白质变体量化方法，当应用于疾病队列时，可以为这些变体在疾病发作、进展和治疗反应中的作用提供有价值的见解。公共卫生相关性：β-2-微球蛋白、半胱氨酸蛋白酶抑制剂C、转铁蛋白、甲状腺素运载蛋白和视黄醇结合蛋白的定量质谱免疫测定的发展将使这些蛋白质及其可能与疾病相关的修饰模式的变化能够进行常规定量监测。有了新的检测方法，我们将更好地研究这些蛋白质在病理过程中的修饰作用，并评估它们作为疾病发作和进展以及对治疗反应的新指标的潜力。