Development of SPR/MS protein array platform

SPR/MS蛋白质芯片平台开发

• 批准号: 6913068
• 负责人: DOBRIN NEDELKOV
• 金额: $ 14.7万
• 依托单位: INTRINSIC BIOPROBES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6913068
• 关键词: affinity labeling; bioengineering /biomedical engineering; biomedical equipment development; biotechnology; blood proteins; blood tests; clinical research; high throughput technology; human subject; immunologic assay /test; mass spectrometry; protein binding; protein protein interaction; protein quantitation /detection; proteomics; surface plasmon resonance; urinalysis

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）： 该提案的主要技术目标是开发表面等离子共振质谱（SPR/MS）蛋白质阵列平台，利用表面等离子共振（SPR）和MALDI-TOF质谱来检测蛋白质和描述蛋白质-蛋白质相互作用。在第一个可行性/试验阶段，我们将检查 SPR 活性表面上的蛋白质排列、阵列表面上蛋白质的亲和力检索以及蛋白质相互作用的 MALDI-TOF MS 读数。功能活性蛋白质阵列将通过将抗体和蛋白质点样（阵列）和固定到芯片表面来创建。然后，蛋白质阵列将用于从溶液中亲和检索蛋白质，之后将通过 MALDI-TOF 质谱分析阵列，以衡量从蛋白质阵列上的点中亲和捕获蛋白质的 MS 读数的可行性。成功完成这些任务后，我们将进入第二个扩展开发阶段，其中将纳入高分辨率 SPR 检测，并且集成的 SPR/MS 蛋白质阵列平台将用于检测生物体液中的蛋白质和蛋白质-蛋白质相互作用。将采用高分辨率 SPR 阵列仪器来显示阵列上蛋白质相互作用（来自各个点）定量的可行性。 SPR/MS 蛋白质阵列平台的组件以及实验控制和变量之间的接口将得到进一步开发和优化。如果需要，将整合更高性能的微点阵仪，并评估 SPR/MS 蛋白质阵列平台在检测血浆和尿液等生物液体中的蛋白质和蛋白质-蛋白质相互作用方面的性能。这项开发研究的最终结果将是一个蛋白质芯片平台和方法，可用于蛋白质组学研究的各个领域，包括高通量生物标志物分析、蛋白质-蛋白质相互作用、群体筛选工作、蛋白质的治疗监测、疾病机制结构的探索和诊断分析的开发。最终，SPR/MS 蛋白质阵列平台可以使用通过微创样本采集方法获得的样本，实现蛋白质生物标志物的快速、并行和高通量筛选，并将筛选工作推广到临床和诊断实验室。