Two-dimensional polyacrylamide gel electrophoresis (2DGE) can detect thousands of polypeptides, separating them by apparent molecular weight and isoelectric point. It provides a more realistic and global review of cellular genetic expression than any other technique. Computer 2DGE analysis software for a variety of platforms has been developed to deal with what can easily be an unwieldy data management problem. However, these programs require significant user input for gel matching and do not adequately correct for routine gel-to-gel distortions. The best approach for gel matching and spot detection is pixel matching from one gel to another. This procedure is computationally demanding and is currently limited to high-end workstations costing 80,000 dollars and above. This Phase I effort proposes to utilize a pixel matching algorithm and integrate an enhanced image processor that will fully automate spot detection for a PC-based system. This system will perform at the level of the expensive workstations but cost 40-50 percent less. Phase I goals include hardware integration of a new image processor PC card into a Pentium class computer, software development for pixel matching of reference to study gel and preliminary testing of the software to meet expected benchmark processing speeds for 2D plasma protein maps. Full automation and increased performance in a PC-based product will facilitate the use of 2DGE in the clinical setting and in proteome research where the focus is on understanding the order, regulation, and coordination of the human genome.
The proposed project will result in a PC-based system capable of fully automated spot detection for 2D gel analysis. The algorithm used will correct for gel-to-gel variations without significant user input. This affordable system will speed up 2D gel analysis and will allow for wider clinical use of this valuable analytical technique.
