SBIR Phase I: Automated 2D Protein Cell Mapping

SBIR 第一阶段：自动化 2D 蛋白质细胞图谱

• 批准号: 0215304
• 负责人: Jack Syage
• 金额: $ 9.99万
• 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: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0215304&HistoricalAwards=false
• 关键词: SBIR; Phase; Automated; 2D; Protein

0215304SyageThis Small Business Innovation Research Phase I Project proposes to test a method for conductinghigh-throughput, automated analysis of the protein content of cell lines using a novel mass analyzed 2D liquid-phase separation technique. The conventional method of 2D PAGE (Polyacrylamide Gel Electrophoresis) 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. This 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 of cell lines to better understand whole system biological function for improved drug therapy and early detection of disease, such as cancer.

0215304Syage这个小型企业创新研究第一阶段项目提议测试一种使用新型质量分析二维液相分离技术对细胞系蛋白质含量进行高通量、自动分析的方法。 2D PAGE（聚丙烯酰胺凝胶电泳）的传统方法有一些局限性。它是劳动密集型的、耗时的、难以自动化并且通常不易再现。此外，定量，特别是在差异表达实验中，通常很困难并且动态范围有限。所提出的技术提供自动化、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。这些功能对于研究药物-蛋白质相互作用以检测癌症的早期迹象将很有价值。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。所提出的系统基于二维液相蛋白质分离，在一维中使用色谱聚焦（CF），在第二维中使用无孔二氧化硅反相高效液相色谱（NPS-RP HPLC）。通过在线电喷雾电离 (ESI) 质谱 (MS) 实时监测 HPLC 洗脱液，以提供分子量和强度信息。该项目的商业应用是在蛋白质组学领域。预计未来 5 年内该市场将从 7 亿美元增长到 58 亿美元。迫切需要开发细胞系蛋白质分析的自动化方法，以更好地了解整个系统的生物学功能，从而改进药物治疗和早期检测癌症等疾病。