Differences in sialic acid O-acetylation between human urinary and recombinant erythropoietins: a possible mass spectrometric marker for doping control

Drug Testing and Analysis
Christian Reichel

Abstract

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...Continue Reading

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Citations

Nov 11, 2014·Expert Review of Proteomics·Mario ThevisWilhelm Schänzer
Nov 14, 2015·Analytical Chemistry·Raul NicoliJean-Luc Veuthey
Dec 20, 2013·Drug Testing and Analysis·Tiia Kuuranne
Mar 29, 2014·British Journal of Sports Medicine·Christian Reichel
Dec 12, 2017·Mass Spectrometry Reviews·Qiwei ZhangJianjun Li
Sep 6, 2019·British Journal of Clinical Pharmacology·Fernando de MoraRafael Martínez

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