High-throughput proteomics using submicroliter amounts of plasma for comprehensive assessment of the immune status

使用亚微升血浆进行高通量蛋白质组学综合评估免疫状态

基本信息

批准号：
10381719
负责人：
Hanno Steen
金额：
$ 117.82万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-09 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10381719
关键词：
Adolescent Adult Agreement Aliquot Antibodies Antibody titer measurement Autoimmunity Bacterial Infections Biological Assay Biological Markers Blood Plasma Volume Blood Volume Boston C-reactive protein Cells Communicable Diseases Communities Complement Consumption Data Development Disease Elderly Erythrocyte Sedimentation Rate Fibrinogen Frail Elderly Funding Funding Opportunities Future Health Hepatitis B Surface Antigens Hepatitis B Vaccination Hepatitis B Vaccines Human Human body Hypersensitivity Immune Immune response Immune system Immunity Immunoglobulin Domain Immunoglobulins Immunologic Deficiency Syndromes Immunologic Monitoring Immunology Individual Infection Maps Measures Methods Modality Molecular Monitor Mycoses National Institute of Allergy and Infectious Disease Neonatal Newborn Infant Organism Parasitic infection Participant Patients Pediatric Hospitals Persons Plasma Plasma Proteins Population Populations at Risk Preparation Prevention Proteins Proteome Proteomics Regulation Research Research Design Resolution Sampling Serology Set protein Specimen System Systems Biology Term Birth Testing Time Transplantation Vaccine Design Virus Diseases Vulnerable Populations Work age group anti-hepatitis B base clinical biomarkers cohort complement pathway complement system experimental study gene product immune function immunological status immunoregulation improved innovation insight interest liquid chromatography mass spectrometry milliliter nanolitre novel premature preterm newborn procalcitonin response vaccine immunogenicity vaccinology

项目摘要

SUMMARY Often, many different tests are performed to obtain a multiparameter assessment of the innate and adaptive immune status. Such tests often include erythrocyte sedimentation rate, C-reactive protein (CRP), procalcitonin, various complement pathway proteins, total antibody (Ab) isotypes and/or subclasses, and specific Ab titers. All these tests are performed on different analytical platforms requiring a separate aliquot for each test. Thus, even with modest sample consumption per test, there is a substantial cumulative volume required for multiple independent tests. While these sample requirements may not be prohibitive for healthy adolescents and adults, they are particularly challenging in especially vulnerable populations such as premature newborns, and frail and/or elderly individuals. For these populations, a wide range of different tests is often very important, however, samples are often limited. In agreement with the purpose of the funding opportunity “Development of Sample Sparing Assays for Monitoring Immune Responses”, we propose to move four different protein assays onto a common processing and analysis platform, which can be performed on the same submicroliter aliquot of plasma thereby significantly reducing the sample volume requirements. The four protein assays of interest include i) an expanded panel of complement components, ii) CRP, iii) antibody isotypes and subclasses including a novel and innovative immunoglobulin domain-resolved map of the IgOme-related protein repertoire (aka IgOme), and iv) the classical plasma proteome, i.e. ~400 well described proteins many of which of clear immunological function. The quantitative information from all these assays will provide us with an exquisite map of the immune status, a hypothesis that we will be tested by correlating our findings with the anti-hepatitis B titer measured before and after hepatitis B vaccination in >500 newborns. Samples will be provided by Boston Children's Hospital's NIAID-funded HIPC (Human Immunology Project Consortium), which will significantly increase the impact of this cohort beyond its original scope. Taking the request of this funding opportunity for sample sparing to a different level, we will advance our plasma proteomics pipeline to quadruple the throughput to at least 96 samples/day and to quarter the plasma volume requirements to <100 nanoliter. Thus, in the future, 1 microliter of plasma will be sufficient for tens of temporally independent LC/MS- based protein assays, resulting in quantitative information for hundreds of proteins such that even a miniscule amount of plasma from the smallest and/or most frail patients will be sufficient to establish an exquisitely detailed and unbiased molecular map of immune status of any person.

概括通常，进行了许多不同的测试以获得先天的多参数评估自适应免疫抑制剂。这样的测试通常包括红细胞沉降率，C反应蛋白（CRP），普罗维替宁，各种补体途径蛋白，总抗体（AB）同种型和/或子类和特定的AB滴度。所有这些测试均在不同的分析平台上进行为每个测试需要一个单独的等分试样。即使每次测试进行适度的样品消耗，是多个独立测试所需的大量累积量。而这些样本对于健康的青少年和成年人，可能不禁止要求，特别是在特别脆弱的人口（例如早产新生儿）和脆弱和/或较早的人群中具有挑战性个人。对于这些人群，各种不同的测试通常非常重要，但是样品通常受到限制。与资金机会的目的一致，“开发样本保证测定法监测免疫反应”，我们建议将四种不同的蛋白质测定移至一个共同处理和分析平台，可以在等离子体的同一亚微光等分试样执行从而大大减少样本量的要求。感兴趣的四种蛋白质测定包括i）扩展的完成组件，ii）CRP，iii）抗体同种型和亚类包括一张新型和创新的免疫球蛋白结构域分辨的蛋白质图曲目（又名Igome）和iv）经典的等离子体蛋白质组，即〜400个描述良好的蛋白质哪些明确的免疫功能。所有这些测定法的定量信息将提供我们拥有免疫状态的独家地图，这是我们将通过关联我们的测试的假设抗肝炎B滴度的发现> 500> 500 新生儿。样品将由波士顿儿童医院的NIAID资助HIPC提供（人类免疫学项目联盟），这将大大增加该队列的影响原始范围。我们请求这个资金机会以将样本保留到不同的水平，我们将推进我们的血浆蛋白质组学管道，使吞吐量四倍至少96个样本/天并四分之一的等离子体量要求到<100纳米仪。将来，1微级等离子体足以足以足以进行数十个暂时独立的LC/MS- 基于蛋白质的蛋白质测定，从而获得数百种蛋白质的定量信息来自最小和/或大多数脆弱患者的小血浆足以建立任何人的免疫状态的精确详细且无偏的分子图。