Ultra-Sensitive Multiplexed Quantification of Proteins Secreted from Single-Cells - Resubmission 01

对单细胞分泌的蛋白质进行超灵敏多重定量 - 重新提交 01

基本信息

批准号：
10326373
负责人：
Savas Tay
金额：
$ 39.53万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10326373
关键词：
ART protein Address Antibodies Area Binding Biological Biological Assay Cells Cellular Assay Complement Confusion DNA Data Detection Development Devices Ensure Event Gene Expression Regulation Growth Factor Hormonal Immune Immunity Individual Insulin Investigation Islet Cell Laboratories Life Malignant Neoplasms Measurement Measures Medical Metabolism Methods Microfluidic Microchips Microfluidics Molecular Organism Peptides Population Protein Secretion Proteins Proteomics Reporting Signal Pathway Signal Transduction Signaling Protein Sorting - Cell Movement Spottings Surface System Techniques Technology Testing Time Work amplification detection base biological information processing biological systems cell type chemokine cytokine detection assay detection method digital experimental study graphical user interface hormonal signals microfluidic technology miniaturize new technology operation pancreatic cancer cells pancreatic juice prototype single cell proteins user friendly software user-friendly wound healing

项目摘要

Ultra-Sensitive Multiplexed Quantification of Proteins Secreted from Single-Cells This proposal aims to develop a powerful new technology for ultra-sensitive quantification of multiple proteins secreted from single-cells. Secreted protein signals coordinate cellular and organism-level functions across all biological systems, and their accurate quantification is crucial for understanding biological information processing in general. Unfortunately, signaling studies have been hindered by poor sensitivity and limited multiplexing ability of current proteomic methods, and the confusion of results due to pooling of multiple cell readouts when conducting population-averaged assays. To address the need to quantify single-cell secreted proteins across different signaling systems, we will develop a new microfluidic technology for ultra-sensitive, multiplexed protein quantification at the single-cell level. We will use our wide-ranging expertise in single-cell assay and microfluidic development and our extensive preliminary data to develop a user-friendly system that will measure multiplexed protein signals from a wide range of cell types, in a time-dependent manner. We will validate our method by quantifying signaling time courses of diverse systems including immune cells, cancer secretion, and hormone signaling. This method will, for the first time, provide the means to gather sensitive, multiplexed single-cell secreted protein measurements, and will be indispensable for understanding signaling in immunity, metabolism, development and cancer.

从单细胞分泌的蛋白质的超敏感多路复用定量该建议旨在开发一种强大的新技术，用于对多种蛋白质进行超敏感从单细胞分泌。分泌的蛋白质信号协调细胞和生物水平的功能所有生物系统及其准确的定量对于理解生物学信息至关重要一般处理。不幸的是，信号研究受到敏感性不佳和有限的限制。当前蛋白质组学方法的多路复用能力，以及由于多个单元的聚集而引起的结果混乱进行人口平均测定法时的读数。解决量化单细胞分泌的需求跨不同信号系统的蛋白质，我们将开发一种新的微流体技术，用于超敏感，单细胞水平的多重蛋白质定量。我们将在单细胞中使用广泛的专业知识测定和微流体开发以及我们广泛的初步数据，以开发一个用户友好的系统这将以时间依赖的方式测量来自各种细胞类型的多路复用蛋白信号。我们将通过量化包括免疫细胞在内的不同系统的信号传导时间过程来验证我们的方法，癌症分泌和激素信号传导。这种方法将首次提供收集的手段敏感的，多重的单细胞分泌蛋白质测量值，对于必不可少的了解免疫，代谢，发育和癌症中的信号传导。