SBIR Phase II: Automated 2D Protein Cell Mapping

SBIR 第二阶段：自动二维蛋白质细胞图谱

• 批准号: 0321763
• 负责人: Jack Syage
• 金额: $ 49.88万
• 依托单位: SYAGEN TECHNOLOGY INC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-11-15 至 2006-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0321763&HistoricalAwards=false
• 关键词: SBIR; Phase; II; Automated; 2D

This Small Business Innovation Research Phase I project will develop a method for conducting high-throughput, automated analysis of the protein content of cell lines using a novel mass analyzed two-dimensional liquid-phase separation method. The conventional method of two-dimensional poly-acrylamide gel electrophoresis (2D PAGE) has several limitations ; it is labor intensive, time consuming, difficult to automate and often not readily reproducible. In addition, quantitation, especially in differential expression experiments, is often difficult and limited in dynamic range. The proposed technology provides automated, faster, and more accurate 2D protein maps, and can be used to purify specific proteins and enact protein/peptide digest and sequencing information. These capabilities will prove valuable for studying drug-protein interactions for detecting early signs of cancer. Studies of cancer cell lines can reveal signatures of cancerous cells that can serve as markers for actual diagnosis. The proposed system is based on 2D liquid-phase protein separation using chromatofocusing (CF) in one dimension and non-porous silica, reverse-phase, high-performance liquid chromatography (NPS-RP HPLC) in the second dimension. The HPLC eluent is monitored in real-time by on-line electrospray ionization (ESI) mass spectrometry (MS) to provide molecular weight and intensity information.The commercial application of this project is in the area of proteomics. The proteomics market is forecasted to grow from $ 0.7 billion to $ 5.8 billion over the next 5 years. There is a tremendous need to develop automated methods of protein analysis and peptide analysis of cell lines to better understand global biological function for improved drug therapy and early detection of disease, such as cancer.

这个小企业创新研究第一阶段项目将开发一种方法，用于使用一种新的质量分析二维液相分离方法对细胞系的蛋白质含量进行高通量，自动化分析。二维聚丙烯酰胺凝胶电泳（2D PAGE）的传统方法有几个局限性，它是劳动密集型，耗时，难以自动化，往往不容易重现。此外，定量，特别是在差异表达实验中，通常是困难的，并且在动态范围内受到限制。所提出的技术提供了自动化，更快，更准确的2D蛋白质图谱，可用于纯化特定蛋白质，并制定蛋白质/肽消化和测序信息。这些能力将被证明对研究药物-蛋白质相互作用以检测癌症的早期迹象很有价值。对癌细胞系的研究可以揭示癌细胞的特征，这些特征可以作为实际诊断的标记。所提出的系统是基于二维液相蛋白质分离使用色谱聚焦（CF）在一维和无孔二氧化硅，反相，高效液相色谱（NPS-RP HPLC）在第二维。通过在线电喷雾离子化（ESI）质谱（MS）实时监测HPLC洗脱液，以提供分子量和强度信息。蛋白质组学市场预计将在未来5年内从7亿美元增长到58亿美元。非常需要开发细胞系的蛋白质分析和肽分析的自动化方法，以更好地了解整体生物学功能，用于改进药物治疗和早期检测疾病，如癌症。