Optical imaging of size, charge, mobility and binding of single proteins

单个蛋白质的大小、电荷、迁移率和结合的光学成像

• 批准号: 10687006
• 负责人: SHAOPENG WANG
• 金额: $ 29.45万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687006
• 关键词: Affinity; Antibodies; Binding; Binding Proteins; Biochemical Process; Biological Markers; Biosensor; Buffers; Capillary Electrophoresis; Cells; Charge; Chemistry; Complication; Computer software; Consumption; Coupled; Detection; Diagnosis; Diameter; Disease; Drug Screening; Electrophoresis; Enzyme-Linked Immunosorbent Assay; Frequencies; Heterogeneity; Image; Imaging technology; In Situ; Individual; Industry; Kinetics; Label; Length; Ligand Binding; Ligands; Light; Link; Measurement; Measures; Mechanics; Methods; Molecular; Neoplasm Circulating Cells; Organism; Pharmaceutical Preparations; Phase; Polymers; Process; Property; Protein Analysis; Proteins; Protocols documentation; Research; Sampling; Signal Pathway; Signal Transduction; Slide; Surface; Surface Plasmon Resonance; System; Technology; Testing; Time; Western Blotting; Workload; bioelectronics; cost; density; detection platform; disease diagnosis; electric field; exosome; experimental study; fabrication; flexibility; gel electrophoresis; indium tin oxide; instrument; light scattering; molecular marker; molecular scale; optical imaging; protein biomarkers; protein complex; protein protein interaction; prototype; sensor; single molecule; success; tool

PROJECT SUMMARY Protein analysis is essential to the understanding of molecular scale processes in living systems, the diagnosis of diseases based on molecular biomarkers, and the treatment of diseases with drugs. The basic tasks of protein analysis include detecting a protein, identifying it and determining its interactions with other proteins or molecular ligands. Various technologies have been developed to perform these tasks, but the most indispensable ones are gel and capillary electrophoresis, Western Blot (WB) and enzyme linked immunosorbent assay (ELISA). These technologies separate and identify proteins based on a protein’s charge, size, and specific binding to antibodies. For molecular interaction analysis, surface plasmon resonance and other detection technologies are the current choices. Although ubiquitous in both research labs and industry, these platforms must be combined to provide complete analysis of proteins, which is complicated and time consuming. In addition, they lack single molecule analysis capability required for studying heterogenous processes and for achieving precision diagnosis, especially for low volume samples. The present project aims to develop one detection platform that can perform the key functions of the above technologies with single molecule detection capability. The proposed technology images single proteins without labels, measures the size, charge and mobility of each protein simultaneously, identifies the protein based on its specific binding to antibodies, and quantifies its interactions with other proteins in real time. The team at the Biodesign Center for Bioelectronics and Biosensors, ASU, has carried out substantial experiments to demonstrate the proposed technology. In this R01 project, the team will address remaining technical challenges, build a complete prototype and validate it for single protein analysis on single cells.

项目摘要 蛋白质分析对于理解生命系统中的分子尺度过程、诊断 基于分子生物标志物的疾病诊断和药物治疗。蛋白质的基本任务 分析包括检测蛋白质、鉴定蛋白质并确定其与其它蛋白质或分子的相互作用 配体。已经开发了各种技术来执行这些任务，但最不可或缺的是 凝胶和毛细管电泳、蛋白质印迹（WB）和酶联免疫吸附测定（ELISA）。这些 技术根据蛋白质的电荷、大小和与抗体的特异性结合来分离和鉴定蛋白质。 对于分子间相互作用分析，表面等离子体共振等检测技术是当前研究的热点 选择.尽管这些平台在研究实验室和工业中无处不在，但必须将它们结合起来， 蛋白质的完整分析，这是复杂和耗时的。此外，它们缺乏单分子 研究异质过程和实现精确诊断所需的分析能力， 特别是对于小体积样品。本项目旨在开发一个检测平台， 上述技术的关键功能与单分子检测能力。所提出的技术 对没有标记的单个蛋白质进行成像，同时测量每个蛋白质的大小、电荷和迁移率， 基于蛋白质与抗体的特异性结合来鉴定蛋白质，并量化其与其他蛋白质的相互作用 在真实的时间里。亚利桑那州立大学生物电子学和生物传感器生物设计中心的团队已经进行了 大量的实验来证明所提出的技术。在这个R01项目中，团队将解决 剩余的技术挑战，建立一个完整的原型，并验证它的单一蛋白质分析的单一 细胞