Two-dimensional capillary electrophoresis - final report of the Innovative Project I4215

When analyzing complex protein mixtures,two-dimensional gel electrophoresis is often used. This technique is time-consuming, demands special instrumentation and includes many manual steps. By using capillary-based methods in a two-dimensional combination, smaller sample amounts can be used, the total analysis time is thus shortened and the manual steps are reduced. The first dimension is capillary isoelectric focusing, where proteins are separated according to their pI. The separated and focused proteins are then transferred to the second dimension via an enzymatic column where they are digested into peptides. In the second dimension, these peptides are separated by liquid chromatography coupled to mass spectrometry. In the mass spectrometer, the individual peptides are identified and thereby the original proteins. This two-dimensional combination can both be used for analyzing complex protein mixtures and as a tool to get detailed information about the ricin toxin.