Electrochemically-Enhanced Plasmonic Imaging for Quantitative Proteomics

用于定量蛋白质组学的电化学增强等离子体成像

基本信息

批准号：
8976613
负责人：
Nguyen Ly
金额：
$ 56.09万
依托单位：
BIOSENSING INSTRUMENT, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-15 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8976613
关键词：
Address Algorithms Antibodies Applications Grants Area Arizona Behavior Binding Binding Proteins Biological Assay Biological Models Biosensing Techniques Biosensor Carbonic Anhydrase II Cell Surface Proteins Cell physiology Cells Cultured Cells Data Collection Detection Development Drug Interactions Environment Fluorescence Glycols Glycoproteins Health Image Imaging Techniques Imaging technology In Situ Institutes Kinetics Label Lead Measurement Membrane Glycoproteins Membrane Proteins Methods Monitor Peptides Performance Pharmaceutical Preparations Phase Phosphorylation Phosphotransferases Post-Translational Protein Processing Printing Protein Analysis Protein Dynamics Protein Microchips Proteins Proteomics Reporting Research Resolution Sampling Signal Transduction Small Business Innovation Research Grant Solid Surface Plasmon Resonance System Technology Testing Time Translations United States National Institutes of Health Universities base density electric impedance functional group high throughput analysis imaging system instrument new technology plasmonics protein function protein protein interaction prototype quantitative imaging response small molecule small molecule inhibitor software development success temporal measurement

项目摘要

DESCRIPTION (provided by applicant): We are proposing a technology to help in three key areas of proteomics including (a) recognition of protein interactions, (b) characterization of post translational modifications, and (c) quantitative measurements at high spatial and/or temporal resolution to address the dynamics of protein interactions. Several significant types of protein interactions remain difficult to study with existing technologies. For example, the analysis of membrane protein interactions (mostly glycol proteins) is challenging, because these proteins are not stable outside of their native amphiphilic cellular environment. Analysis of interaction kinetics between small molecules (<500 Da, including a vast majority of metabolites and drugs) and proteins is also lacking, because these molecules are too small for fluorescence labeling, and the binding signals are too weak for label-free detection methods. Similarly problematic is the characterization of protein post-translational modifications, which alter protein behavior due to the attachment of a small functional group after translation. Specifically, we propose an electrochemically-enhanced plasmonic imaging (ECEPI) system to address key needs for quantitative analysis of protein interaction dynamics, including the ability to study membrane protein interactions in their native cellular state, characterization of small molecule interaction and post-translational modifications, measurement of interactions at high spatial and temporal resolution for the study of sub-cellular processes, and performing high-throughput analysis in multi-cellular and microarray formats. The ECEPI system relies upon careful integration of three core technologies: 1) the electrochemical surface plasmon resonance systems that have been successfully commercialized by Biosensing Instrument Inc. (BI) for their unique capabilities and solid performance, 2) a proprietary high resolution distortion-free prism-based surface plasmon resonance (SPR) imaging system currently under development at BI for high-throughput interaction analysis, and 3) a highly sensitive impedance imaging technique invented at Arizona State University. The success of this project will lead to a new instrument that is capable of: 1) Label-free real-time recognition and quantification of protein interaction kinetics; 2) Real-time characterization of post-translational modifications of proteins; 3) Quantitative measurement of small molecule interactions with proteins; 4) In situ quantification of membrane protein (and glycoprotein) interactions in their native cellular environment with cell-based assay; 5) High-resolution analysis of sub-cellular processes and; 6) High-throughput analysis in multi-cellular and microarray formats

描述（由申请人提供）：我们正在提出一种技术来帮助蛋白质组学的三个关键领域，包括（a）蛋白质相互作用的识别，（b）后蛋白质的表征翻译修饰，以及（c）高空间和/或时间分辨率的定量测量，以解决蛋白质相互作用的动态。利用现有技术仍然难以研究几种重要类型的蛋白质相互作用。例如，膜蛋白相互作用（主要是乙二醇蛋白）的分析具有挑战性，因为这些蛋白在其天然两亲性细胞环境之外不稳定。小分子（<500 Da，包括绝大多数代谢物和药物）和蛋白质之间的相互作用动力学分析也很缺乏，因为这些分子太小，无法进行荧光标记，而且结合信号太弱，无法进行无标记检测方法。类似的问题是蛋白质翻译后修饰的表征，由于翻译后小功能基团的附着而改变了蛋白质的行为。具体来说，我们提出了一种电化学增强等离子体成像（ECEPI）系统，以满足蛋白质相互作用动力学定量分析的关键需求，包括研究天然细胞状态下膜蛋白相互作用的能力、小分子相互作用的表征和翻译后修饰，以高空间和时间分辨率测量相互作用，以研究亚细胞过程，并以多细胞和微阵列格式进行高通量分析。 ECEPI 系统依赖于三项核心技术的仔细集成：1) 电化学表面等离子体共振系统，该系统已由 Biosensing Instrument Inc. (BI) 因其独特的功能和可靠的性能而成功商业化，2) 专有的高分辨率无失真BI 目前正在开发基于棱镜的表面等离子共振 (SPR) 成像系统，用于高通量相互作用分析；3) 亚利桑那州立大学发明的高灵敏度阻抗成像技术。该项目的成功将带来一种新仪器，它能够：1) 蛋白质相互作用动力学的无标记实时识别和定量； 2）蛋白质翻译后修饰的实时表征； 3）小分子与蛋白质相互作用的定量测量； 4) 通过基于细胞的测定对天然细胞环境中的膜蛋白（和糖蛋白）相互作用进行原位定量； 5) 亚细胞过程的高分辨率分析； 6) 多细胞和微阵列格式的高通量分析