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Capillary electrophoresis time‐of‐flight mass spectrometry for a confident elucidation of a glycopeptide map of recombinant human erythropoietin
Author(s) -
Giménez Estela,
RamosHernan Raquel,
Benavente Fernando,
Barbosa José,
SanzNebot Victoria
Publication year - 2011
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.5114
Subject(s) - chemistry , glycopeptide , chromatography , capillary electrophoresis , mass spectrometry , glycan , time of flight mass spectrometry , recombinant dna , fragmentation (computing) , glycoprotein , analytical chemistry (journal) , biochemistry , organic chemistry , ion , computer science , ionization , operating system , gene , antibiotics
Capillary electrophoresis coupled to orthogonal accelerated time‐of‐flight mass spectrometry (CE/TOFMS) was used for the analysis of O ‐ and N ‐glycopeptides of recombinant human erythropoietin (rhEPO). O 126 and N 83 with a tetraantennary complex type glycan (N 83 ‐4Ant) were selected as glycopeptide models to develop an optimum CE/TOFMS methodology capable of detecting and characterizing the wide variety of glycopeptides present in the glycoprotein digest. Glycopeptide adsorption in the inner surface of the fused‐silica capillary was prevented after using a capillary conditioning of 1 M HAc between runs. On the other hand, different acidic conditions in the sheath liquid (SL) and in the background electrolyte (BGE) were tested with the aim of studying their influence in glycopeptide fragmentation. Finally, the fragmentor voltage value of the TOF‐MS instrument was optimized to avoid the involuntary fragmentation of the native glycopeptides. Hence, the established method may be regarded as an excellent starting point to obtain reliable glycopeptide maps of complex glycoproteins such as rhEPO by CE/TOFMS. Copyright © 2011 John Wiley & Sons, Ltd.