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

• 批准号: 8524025
• 负责人: Nguyen Ly
• 金额: $ 35万
• 依托单位: BIOSENSING INSTRUMENT, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8524025
• 关键词: Address; Algorithms; Antibodies; Applications Grants; Area; Arizona; Behavior; Binding; 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 Binding; 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; inhibitor/antagonist; instrument; new technology; plasmonics; protein function; protein protein interaction; prototype; response; small molecule; software development; success

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）：我们提出了一项技术，以帮助蛋白质组学的三个关键领域，包括（a）识别蛋白质相互作用，（b）邮政的表征 转化修饰，以及（c）在高空间和/或时间分辨率下定量测量，以解决蛋白质相互作用的动力学。现有技术的几种重要类型的蛋白质相互作用仍然难以研究。例如，膜蛋白相互作用（主要是乙二醇蛋白）的分析是具有挑战性的，因为这些蛋白质在其天然两亲性细胞环境之外并不稳定。还缺乏小分子之间的相互作用动力学分析（包括绝大多数代谢产物和药物）和蛋白质之间的相互作用动力学，因为这些分子对于荧光标记而言太小，并且结合信号太弱，对于无标记的检测方法而言太弱。同样有问题的是蛋白质后翻译修饰的表征，这会改变翻译后小型功能组的附着蛋白质行为。具体而言，我们提出了一种电化学增强的等离子成像（ECEPI）系统，以解决对蛋白质相互作用动态进行定量分析的关键需求，包括研究其天然细胞状态中膜蛋白相互作用的能力，对小分子相互作用的表征和在高分子后的相互作用，在高跨度的相互作用中，测量跨性别的相互作用，测量跨性别的次数，研究，测量了跨性别的互动，研究了跨性别的相互作用，研究了跨性别的相互作用，研究了对跨分子相互作用的相互作用，并进行了互动，研究了对膜细胞状态的相互作用，并进行了互动，研究了对膜细胞状态的相互作用，并进行了互动的互动。多细胞和微阵列格式的分析。 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成像技术在亚利桑那州立大学发明。该项目的成功将导致一种能够：1）的新工具 无标记的实时识别和蛋白质相互作用动力学的定量； 2）蛋白质翻译后修饰的实时表征； 3）定量测量小分子与蛋白质的相互作用； 4）原位定量膜蛋白（和糖蛋白）在其天然细胞环境中与基于细胞的测定的相互作用； 5）对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