Premium
Capillary electrophoresis and matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry for analysis of the novel erythropoiesis‐stimulating protein (NESP)
Author(s) -
SanzNebot Victoria,
Benavente Fernando,
Giménez Estela,
Barbosa José
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200410092
Subject(s) - chemistry , capillary electrophoresis , mass spectrometry , time of flight mass spectrometry , chromatography , matrix assisted laser desorption/ionization , ionization , analytical chemistry (journal) , erythropoietin , resolution (logic) , molecular mass , mass spectrum , matrix (chemical analysis) , desorption , erythropoiesis , ion , biochemistry , medicine , enzyme , organic chemistry , adsorption , artificial intelligence , computer science , anemia
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.