Dynamic Isoelectric Focusing for Proteomics

Abstract

Dynamic isoelectric focusing is a new technique that is related to capillary isoelectric focusing but uses additional high-voltage power supplies to provide control over the shape of the electric field within the capillary. Manipulation of the electric field changes the pH gradient, enabling both the location and width of the focused protein bands to be controlled. The proteins can be migrated to a designated sampling point while remaining focused, where they can be collected for further analysis. This ability to collect and isolate the protein bands while maintaining a high peak capacity demonstrates that dynamic isoelectric focusing has great potential as a first dimension in a multidimensional separation system. Dynamic isoelectric focusing can achieve a peak capacity of over 1000, as shown by both mass spectrometry analysis and direct imaging