title: Screening of Small Intact Proteins by Capillary Electrophoresis Electrospray Ionization-Mass Spectrometry (CE-ESI-MS).
abstract: 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.
authors.affiliation: Department of Chemistry, Aalen University, Beethovenstraβe 1, Aalen, D-73430, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Neuberger
authors.affiliation: Solvias AG, Kaiseraugst, Switzerland.
authors.forename: Angelina
authors.initials: A
authors.lastname: Rafai
authors.affiliation: Department of Chemistry, Aalen University, Beethovenstraβe 1, Aalen, D-73430, Germany. christian.neusuess@htw-aalen.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Neusüß
mesh_terms: D020099:Coated Materials, Biocompatible; D019075:Electrophoresis, Capillary / Q000379:methods*; D004921:Erythropoietin / Q000302:isolation & purification*; D006801:Humans; D011994:Recombinant Proteins / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D020099:Coated Materials, Biocompatible; C554270:EPO protein, human; D011994:Recombinant Proteins; D004921:Erythropoietin
keywords: Capillary electrophoresis; Coating; Electrospray ionization; Intact protein; Mass spectrometry