Advances in two-dimensional gel matching technology

Abstract

For many years, two-dimensional gel electrophoresis has been the method of choice for the investigation of complex mixtures of proteins. Although there are a number of emerging technologies that can be applied to proteomics, none can yet yield routinely the breadth of information available from two-dimensional gels. To be able to obtain instant information regarding molecular mass and pI, as well as to highlight quickly the expression changes or unique proteins across a gel series requires sophisticated and powerful image analysis software. The range of software products offered by Nonlinear Dynamics covers all levels of user application and throughput, from the user-guided Phoretix two-dimensional approach, when working with a small number of gels, to the automatic processing of large numbers of gels with minimal user intervention with Progenesis. Integration of the analysis software with powerful database components allows advanced gel comparisons and data mining to be performed with statistical verification of the results. Spot pick lists can be quickly created and automatically linked to a number of commercially available spot picking robots further increasing the support for proteomics research. The importance of image analysis for accurate, reliable and meaningful results will be discussed. Recent advances in development, with particular attention placed on the impact of noise contamination within gels, are illustrated and how the Progenesis product from Nonlinear Dynamics can be utilized to get the most from two-dimensional gel electrophoresis is shown.