Development of SPR/MS protein array platform

SPR/MS蛋白芯片平台开发

• 批准号: 7244363
• 负责人: DOBRIN NEDELKOV
• 金额: $ 36.45万
• 依托单位: INTRINSIC BIOPROBES, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7244363
• 关键词: Affinity; Antibodies; Binding; Binding Proteins; Biological; Biological Markers; Biosensor; Blood; Caliber; Cell surface; Chemistry; Clinical; Detection; Development; Diagnostic; Dimensions; Disease; Immobilization; Individual; Invasive; Laboratories; Lasers; Ligands; Liquid substance; MALDI-TOF Mass Spectrometry; Mass Spectrum Analysis; Methods; Modeling; Monitor; Performance; Phase; Plasma; Population; Preparation; Process; Protein Array; Protein Array Analysis; Protein Microchips; Proteins; Proteomics; Research; Resolution; Retrieval; Route; Sampling; Screening procedure; Signal Transduction; Solutions; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spottings; Structure; Surface; Surface Plasmon Resonance; Therapeutic; Urine; Width; Work; assay development; density; high throughput screening; instrument; mass spectrometer; protein protein interaction; research study; response; sample collection; sensor; stem

The main technical objective of this proposal is to develop Surface Plasmon Resonance Mass Spectrometry (SPR/MS) protein array platform that utilizes Surface Plasmon Resonance (SPR) and MALDI-TOF mass spectrometry for detection of proteins and delineation of protein-protein interactions. In the first feasibility/pilot phase, we will examine the protein arraying to an SPR-active surface, affinity retrieval of proteins on the array surface, and MALDI-TOF MS readout of the protein interactions. Functionally-active protein array will be created via spotting (arraying) and immobilization of antibodies and proteins onto a chip surface. The protein array will then be used for affinity-retrieval of proteins from solution, after which the array will be analyzed via MALDI-TOF mass spectrometry to gauge the feasibility of the MS readout of the affinity-captured proteins from the spots on the protein array. Upon the successful completion of these tasks, we will move into the second, expanded development phase, where high-resolution SPR detection will be incorporated and the integrated SPR/MS protein array platform will be used for detection of proteins and protein-protein interactions from biological fluids. A high resolution SPR array instrument will be employed to show the feasibility of quantification of the protein interactions (from individual spots) on the array. The interface between the components of the SPR/MS protein array platform, and the experiment controls and variables, will be further developed and optimized. If needed, a higher-performance microarrayer will be incorporated, and the performance of the SPR/MS protein array platform in detection of proteins and protein-protein interactions from biological fluids such as plasma and urine will be evaluated. The final result of this developmental research will be a protein chip platform and methods that can be employed into various lines of proteomics research, including high-throughput biomarker analysis, protein-protein interactions, population screening efforts, therapeutic monitoring of proteins, exploration of disease mechanism structures, and diagnostic assays development. Ultimately, the SPR/MS protein array platform could enable rapid, parallel, and high-throughput screening of protein biomarkers, using samples obtained through minimally invasive sample collection methods, propagating the screening efforts into the clinical and diagnostic laboratories.

该建议的主要技术目标是发展表面等离子体共振质谱法 （SPR/MS）使用表面等离子体共振（SPR）和MALDI-TOF质量的蛋白阵列平台 用于检测蛋白质和蛋白质蛋白相互作用的描述的光谱法。在第一个可行性/飞行员中 阶段，我们将检查蛋白质阵列到SPR活性表面，亲和力在蛋白质上的亲和力检索 阵列表面和蛋白质相互作用的MALDI-TOF MS读数。功能活性蛋白阵列将 通过斑点（阵列）以及将抗体和蛋白质固定在芯片表面上创建。这 然后 通过MALDI-TOF质谱分析以评估MS读数的可行性 来自蛋白质阵列斑点的亲和捕获蛋白。成功完成这些任务后， 我们将进入第二个扩展的发展阶段，高分辨率SPR检测将是 Incomporated和集成的SPR/MS蛋白质阵列平台将用于检测蛋白质和 来自生物液的蛋白质蛋白质相互作用。高分辨率SPR阵列仪器将用于 显示阵列上蛋白质相互作用（来自单个斑点）的可行性。这 SPR/MS蛋白阵列平台的组件之间的接口，实验控制和 变量将进一步开发和优化。如果需要，更高绩效的微阵列将是 并入，SPR/MS蛋白阵列平台的性能在检测蛋白质和 将评估来自血浆和尿液等生物液的蛋白质 - 蛋白质相互作用。最终结果 在这项发展研究中，将是一个蛋白质芯片平台和可以用于各种的方法 蛋白质组学研究线，包括高通量生物标志物分析，蛋白质 - 蛋白质相互作用， 人口筛查工作，蛋白质的治疗监测，探索疾病机制结构， 和诊断测定开发。最终，SPR/MS蛋白质阵列平台可以快速， 使用最少获得的样品对蛋白质生物标志物的平行和高通量筛选 侵入性样本收集方法，将筛查工作传播到临床和诊断中 实验室。