Capillary electrophoresis (CE) has been shown to be a suitable separation technique for complex samples. Combined with electrospray ionization-mass spectrometry (ESI-MS), it is a powerful tool offering the opportunity of high selectivity and sensitivity combined with the possibility to identify and characterize intact proteins. In this protocol, we demonstrate a screening method for intact proteins based on capillary zone electrophoresis (CZE) separation coupled with online mass spectrometric detection. In order to avoid protein-wall interactions, a neutral coated capillary is used to create a universal method for proteins with both low and high electrophoretic mobilities. In addition, we show the successful validation and application of this screening method for a set of eight standard proteins and the glycoprotein erythropoietin.
Keywords: Capillary electrophoresis; Coating; Electrospray ionization; Intact protein; Mass spectrometry.