Automated 2D Protein Cell Mapping

This Small Business Innovation Research Phase I Project proposes to test a method for conducting high-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