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（由申请人提供）：该阶段项目的主要目的是为β-2-微球蛋白，胱抑素C，转铁蛋白，经硫代蛋白和视黄醇结合蛋白开发定量质谱免疫测定。最近，通过定性质谱免疫测定1,000个健康队列中测定了这五种蛋白质，检测（不提供浓度信息）多种蛋白质变体。由于基于三明治的免疫测定（例如ELISA）无法明确量化微妙的蛋白质修饰，因此在此项目中，我们建议为这些蛋白质及其同工型开发定量的质谱免疫测定。将仔细选择内部参考标准，并将进行逐步的标准曲线定量质谱质谱法开发。将确定定量质谱免疫测定的精度（测定内和测定），线性，灵敏度和特异性。作为最终验证步骤，将针对现有ELISA分析进行评估和基准测试定量的MSIA分析。这里提出的定量质谱免疫测定将提供量化蛋白质变异的独特而直接的方法，当应用于疾病同类群时，可以为这些变体在疾病，进展和对治疗的反应中的作用提供宝贵的见解。公共卫生相关性：开发用于β-2-微球蛋白，半胱氨酸蛋白酶C，转铁蛋白，经硫代蛋白和视黄醇结合蛋白的定量质谱免疫测定，将实现对这些蛋白质的常规定量监测及其修饰模式的变化。随着新的测定，我们将有能力研究这些蛋白质在病理过程中修饰的影响，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。