Development of a rapid glycosylation profiling system using mass spec data

使用质谱数据开发快速糖基化分析系统

• 批准号: 7695559
• 负责人: HEATHER R DESAIRE
• 金额: $ 29万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7695559
• 关键词: Academic Medical Centers; Address; Algorithms; Antibodies; Biochemical; Biochemistry; Biological Markers; Collaborations; Complex; Computer software; Data; Data Analyses; Development; Disease; Epitopes; Eye; Future; Glycopeptides; Glycoproteins; Goals; HIV; HIV envelope protein; HIV vaccine; HIV-1; Hand; Health; Heel; Human; Internet; Lead; Left; Literature; Manuals; Methods; Peptides; Pharmacologic Substance; Polysaccharides; Positioning Attribute; Process; Protein Analysis; Proteins; Research; Research Personnel; Resolution; Safety; Sampling; Screening procedure; Site; Software Design; Surface; System; Technology; Thinking; Time; Vaccines; Viral; Virus; Work; base; data acquisition; design; env Gene Products; env Glycoproteins; experience; glycosylation; high throughput screening; improved; innovation; neutralizing antibody; programs; protein function; public health relevance; research study; skills; tool; vaccine candidate; vaccine development

DESCRIPTION (provided by applicant): The ultimate goal of this proposal is to develop a fully-automated approach for analysis of glycopeptides and to apply that approach to the analysis of several different HIV-Env glycoproteins. This work would be accomplished by completing three specific aims: AIM 1: Using MS data as the input, develop an algorithm that rapidly identifies glycopeptide compositions for singly glycosylated glycopeptides. AIM 2: Develop an algorithm that characterizes multiply glycosylated glycopeptides. AIM 3: Use the tools in AIMS 1 and 2 to screen for accessible and genetically conserved points on the HIV Env protein. The bulk of the work, Aims 1 and 2, would be completed by synergistically incorporating expertise in mass spectral (MS) data acquisition and analysis with expertise in software design/development. This work can broadly impact human health research because glycopeptide analysis is an enabling bioanalytical technology that can be used to screen for biomarkers of disease or disease state; it can be used to help verify the safety and consistency of glycoprotein-based pharmaceuticals; and it can be used to analyze glycopeptides on HIV Envelope proteins, a major target for HIV vaccine development. After completion of the automated glycosylation profiling system (Aims 1 and 2) this system would be used to identify the glycopeptide composition on a variety of HIV- Env vaccine candidates generated in collaboration with Dr. Barton F. Haynes at Duke University Medical Center. The method for this approach includes preparing tryptic digests of each of the glycoproteins, conducing HPLC-MS and LC-MS/MS analyses on the digested products, and interpreting the results using the developed glycopeptide analysis software. After completion of this work, the glycopeptide profiles of the Env proteins would provide information about epitope accessibility on Env. This is relevant to human health because identifying epitopes on the protein that are consistently exposed across HIV-1 clades is a first step in designing an HIV vaccine that mimics these epitopes and elicits broadly neutralizing antibodies to HIV-1. PUBLIC HEALTH RELEVANCE: We aim to develop tools for the rapid analysis of glycopeptides. Glycopeptides are components of the HIV Env protein, and their analysis could be used to identify Achilles Heels (regions which are consistently exposed and thus vulnerable to antibodies) on the surface of the virus.

描述(由申请人提供)：这项提案的最终目标是开发一种全自动化的糖肽分析方法，并将该方法应用于几种不同的HIV-Env糖蛋白的分析。这项工作将通过完成三个特定的目标来完成：目标1：利用MS数据作为输入，开发一种快速识别单糖化糖肽的糖肽组成的算法。目的2：开发一种表征多个糖基化糖肽的算法。目标3：使用AIMS 1和AIMS 2中的工具来筛选HIV环境蛋白上可访问的和遗传上保守的点。目标1和目标2的大部分工作将通过将质谱学(MS)数据采集和分析方面的专门知识与软件设计/开发方面的专门知识协同结合起来完成。这项工作可以广泛影响人类健康研究，因为糖肽分析是一种生物分析技术，可用于筛选疾病或疾病状态的生物标记物；它可用于帮助验证以糖蛋白为基础的药物的安全性和一致性；它可用于分析HIV包膜蛋白上的糖肽，HIV包膜蛋白是HIV疫苗开发的主要目标。在自动糖基化分析系统(AIMS 1和2)完成后，该系统将用于识别与杜克大学医学中心Barton F.Haynes博士合作产生的各种HIV-Env候选疫苗上的糖肽组成。该方法包括制备每种糖蛋白的胰酶消化液，对消化产物进行高效液相-质谱仪和液质联用仪分析，并使用开发的糖肽分析软件解释结果。在这项工作完成后，环境蛋白的糖肽图谱将提供关于环境蛋白表位可及性的信息。这与人类健康有关，因为识别蛋白质上持续暴露于HIV-1分支的表位是设计模仿这些表位并引发针对HIV-1的广泛中和抗体的HIV疫苗的第一步。公共卫生相关性：我们的目标是开发快速分析糖肽的工具。糖肽是HIV Env蛋白的组成部分，它们的分析可用于识别病毒表面的跟腱(持续暴露于抗体前的区域)。