title: Capillary Zone Electrophoresis-Mass Spectrometry of Intact Proteins.
abstract: Capillary electrophoresis (CE) coupled with mass spectrometry (MS) has proven to be a powerful analytical tool for the characterization of intact proteins. It combines the high separation efficiency, short analysis time, and versatility of CE with the mass selectivity and sensitivity offered by MS detection. This chapter focuses on important practical considerations when applying CE-MS for the analysis of intact proteins. Technological aspects with respect to the use of CE-MS interfaces and application of noncovalent capillary coatings preventing protein adsorption are treated. Critical factors for successful protein analysis are discussed and four typical CE-MS systems are described demonstrating the characterization of different types of intact proteins by CE-MS. These methodologies comprise the use of sheath-liquid and sheathless CE-MS interfaces, and various types of noncovalent capillary coatings allowing efficient and reproducible protein separations. The discussion includes the analysis of lysozyme-drug conjugates and the therapeutic proteins human growth hormone, human interferon-β-1a, and human erythropoietin.
authors.affiliation: Division of BioAnalytical Chemistry, VU University, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Elena
authors.initials: E
authors.lastname: Domínguez-Vega
authors.affiliation: Division of BioAnalytical Chemistry, VU University, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Rob
authors.initials: R
authors.lastname: Haselberg
authors.affiliation: Division of BioAnalytical Chemistry, VU University, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands. g.w.somsen@vu.nl.
authors.forename: Govert W
authors.initials: GW
authors.lastname: Somsen
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D004921:Erythropoietin / Q000032:analysis; D013006:Growth Hormone / Q000032:analysis; D006801:Humans; D000068556:Interferon beta-1a / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D010446:Peptide Fragments / Q000032:analysis; D011506:Proteins / Q000032:analysis*
chemical_list: C554270:EPO protein, human; D010446:Peptide Fragments; D011506:Proteins; D004921:Erythropoietin; C032506:somatotropin (1-134); D013006:Growth Hormone; D000068556:Interferon beta-1a
keywords: (Non-)covalent coatings; Biopharmaceuticals; Capillary electrophoresis; Electrospray ionization; Mass spectrometry; Proteins; Sheathless interface; sheath-liquid interface