Development of SPR/MS protein array platform

SPR/MS蛋白质芯片平台开发

• 批准号: 7238078
• 负责人: DOBRIN NEDELKOV
• 金额: $ 50.53万
• 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/7238078
• 关键词: Development; SPR; MS; protein; array

DESCRIPTION: State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of the project. Describe concisely the research design and methods for achieving these goals. Avoid summaries of past accomplishments and the use of the first person. This abstract is meant to serve as a succinct and accurate description of the proposed work when separated from the application. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE PROVIDED. 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. PERFORMANCE SITE(S) (organization, city, state) Intrinsic Bioprobes Inc., Tempe, AZ KEY PERSONNEL. See instructions. Use continuation pages as neededto provide the required information in the format shown below. Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name Organization Role on Project Nedelkov, Dobrin Intrinsic Bioprobes Inc. PI Nelson, Randall W. Intrinsic Bioprobes Inc. Co-Pi Disclosure Permission Statement. Applicable to SBIR/STTR Only. Seeinstructions. 53 Yes l~l No PHS 398 (Rev. 05/01) Page_2 Form Page 2 Principal Investigator/Program Director (Last, first, middle): NedelkoV, Dobrin The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT

说明：说明申请的广泛、长期目标和具体目标，并参考项目的健康相关性。描述 简要介绍了实现这些目标的研究设计和方法。避免总结过去的成就和使用第一人称。这个抽象 是为了作为一个简洁和准确的描述拟议的工作时，从申请分开。如果申请得到资助， 这样的描述将成为公开信息。因此，不包括专有/机密信息。不要超过空间 提供了 本提案的主要技术目标是开发表面等离子体共振质谱 (SPR/MS）蛋白质阵列平台，其利用表面等离子体共振（SPR）和MALDI-TOF质谱 用于检测蛋白质和描绘蛋白质-蛋白质相互作用的光谱法。在第一个可行性/试点中 阶段，我们将研究蛋白质阵列到SPR活性表面，亲和修复蛋白质上的蛋白质， 阵列表面，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（排列）和固定到芯片表面上来产生。的 然后，蛋白质阵列将用于从溶液中亲和回收蛋白质，之后，将该阵列 通过MALDI-TOF质谱分析，以衡量MS读出的可行性。 从蛋白质阵列上的点亲和捕获的蛋白质。在圆满完成这些任务后， 我们将进入第二个扩展的开发阶段，在这个阶段，高分辨率的SPR检测将是 整合的SPR/MS蛋白质阵列平台将用于检测蛋白质， 蛋白质与蛋白质之间的相互作用。将采用高分辨率SPR阵列仪器， 显示了在阵列上定量蛋白质相互作用（来自单个点）的可行性。的 SPR/MS蛋白质阵列平台组件与实验对照之间的接口， 变量，将进一步开发和优化。如果需要的话，一个更高性能的微阵列将是 结合，和SPR/MS蛋白质阵列平台在蛋白质检测中的性能， 将评价来自生物流体如血浆和尿的蛋白质-蛋白质相互作用。最终结果 这项发展研究的一部分将是一个蛋白质芯片平台和方法，可以用于各种 蛋白质组学研究，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， 和诊断分析开发。最终，SPR/MS蛋白质阵列平台可以实现快速、 平行和高通量筛选蛋白质生物标志物，使用通过最低限度地 侵入性样本采集方法，将筛查工作推广到临床和诊断领域， laboratories. 履约地点（组织、城市、州） Intrinsic Bioprobes Inc.，滕佩，亚利桑那州 关键人员。参见说明。根据需要使用续页，以下列格式提供所需信息。 从首席研究员开始。按字母顺序列出所有其他关键人员，姓在前。 名称组织在项目中的角色 Nedelkov，Dobrin Intrinsic Bioprobes Inc. Pi 作者：纳尔逊Intrinsic Bioprobes Inc.共同主持 披露许可声明。仅适用于SBIR/STTR。请参阅说明。53是l~l否 PHS 398（修订版05/01）第2页表格第2页 主要研究者/项目负责人（最后一名、第一名、中间名）：Nedelkov、Dobrin 主要研究者/项目负责人的姓名必须在打印页和续页的顶部提供。 研究资助