title: Capillary electrophoresis and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry for analysis of the novel erythropoiesis-stimulating protein (NESP).
abstract: NESP (novel erythropoiesis-stimulating protein) is a recently approved hyperglycosylated analogue of human erythropoietin (EPO) with a long-lasting effect. In this work, the capillary electrophoresis (CE) methodology proposed by the European Pharmacopoeia for the separation of EPO glycoforms has been modified for the separation of NESP glycoforms. Optimization of pH of the separation electrolyte has been fundamental in order to achieve baseline resolution of seven peaks corresponding to NESP glycoforms. Intact NESP has also been characterized by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). An accurate approximation to an average molecular mass of the NESP molecule has been obtained, taking into account the strong influence of laser intensity upon the MALDI-TOF mass spectra found.
authors.affiliation: Department of Analytical Chemistry, University of Barcelona, Barcelona, Spain. vsanz@ub.edu
authors.forename: Victoria
authors.initials: V
authors.lastname: Sanz-Nebot
authors.forename: Fernando
authors.initials: F
authors.lastname: Benavente
authors.forename: Estela
authors.initials: E
authors.lastname: Giménez
authors.forename: José
authors.initials: J
authors.lastname: Barbosa
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D004921:Erythropoietin / Q000032:analysis*; D006397:Hematinics; D011994:Recombinant Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D006397:Hematinics; D011994:Recombinant Proteins; D004921:Erythropoietin