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Capillary electrophoresis and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry for analysis of the novel erythropoiesis-stimulating protein (NESP)

✍ Scribed by Victoria Sanz-Nebot; Fernando Benavente; Estela Giménez; José Barbosa

Publisher
John Wiley and Sons
Year
2005
Tongue
English
Weight
123 KB
Volume
26
Category
Article
ISSN
0173-0835

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✦ Synopsis

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.

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