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Differences in sialic acid O‐acetylation between human urinary and recombinant erythropoietins: a possible mass spectrometric marker for doping control
Author(s) -
Reichel Christian
Publication year - 2013
Publication title -
drug testing and analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.065
H-Index - 54
eISSN - 1942-7611
pISSN - 1942-7603
DOI - 10.1002/dta.1563
Subject(s) - chemistry , acetylation , glycan , glycopeptide , sialic acid , glycosylation , recombinant dna , biochemistry , mass spectrometry , erythropoietin , chromatography , glycoprotein , endocrinology , biology , gene , antibiotics
Development of a mass spectrometric method for the unambiguous detection of doping with recombinant human erythropoietins (rhEPO) has been attempted for many years. Unfortunately, progress in this field was hampered by the unavailability of highly purified human endogenous EPOs (urinary[uhEPO], serum/plasma EPO) – a prerequisite for generating detailed mass spectrometric glycosylation data necessary for revealing significant differences between uhEPO and rhEPOs. The paper presents the worldwide first analytical data on purified human urinary EPO generated with a high resolution high accuracy mass spectrometer (LTQ‐Orbitrap). The focus is on the tryptic O‐glycopeptide (E117‐R131) and its degree of sialic acid O‐acetylation. Data are compared with results obtained from 40 rhEPO pharmaceuticals. It could be demonstrated that the O‐glycopeptide of uhEPO ( ca 100 IU) contains only trace amounts of mono‐acetylated mono‐and di‐sialylated O‐glycans but no other O‐acetylated structures and in this respect significantly differs from all rhEPOs. Moreover, Dynepo – a rhEPO previously thought to be not O‐acetylated – also contains small amounts of O‐acetylations within the O‐glycan structure. The results might be useful for anti‐doping purposes as well as the development of EPO pharmaceuticals with closer structural similarity to the endogenous hormone. Copyright © 2013 John Wiley & Sons, Ltd.