Simone König
University of Münster, Germany
Title: Reproducibility in 2D- gel-based proteome analysis: External and internal standardisation with Comparative 2D Fluorescence Gel Electrophoresis CoFGE
Biography
Biography: Simone König
Abstract
A major problem for the acceptance of two-dimensional (2D) protein gel electrophoresis in proteome expression analysis has been the little comparability of gel images among different laboratories or even within the same working group. Now, the generation of a reference grid of marker proteins in parallel to the separation of the analyte allows the correction of protein coordinates both in x (protein isoelectric point)- and y (molecular weight)-direction. This is achieved by use of multiple fluorescent dyes for standards and samples on the same gel. A net of ~100 marker nodes spans the gel and provides reference points for triangulation. The deviation from mean is improved by an order of magnitude. The technology termed Comparative 2D Fluorescence Gel Electrophoresis (CoFGE) can be carried out in both vertical and horizontal electrophoresis instrumentation with the latter being much more comfortable. With CoFGE, archiving of coordinates of proteins identified by mass spectrometry in databases has become reality and a Repository of Gel-Separated Proteins (ReGeSeP) was generated for storing and searching protein IDs, coordinates and proteome gel images. The technology was protected (Patent application EP11167383.6, May 25, 2011), licensed, and awarded with the WWU Transfer Prize 2013/2014.