Norwegian Doping Control Laboratory, Oslo University Hospital, 0586 Oslo, Norway.
Drug Test Anal. 2011 May;3(5):291-9. doi: 10.1002/dta.270. Epub 2011 Mar 9.
Erythropoietin (EPO) has been misused in sports for many years due to its performance-enhancing effect. In the last decade, detection of abuse has been possible with isoelectric focusing (IEF) based on the different isoform profiles of endogenous and recombinant EPO. The release of new EPOs on the market, such as the recombinant erythropoietin epoetin delta (Dynepo™) and the chemically modified EPO, CERA (Mircera™) potentially represents analytical challenges to the fight against doping. This study set out to investigate the possibility of and the time window for detecting the administration of a single dose of Dynepo™ and CERA. Our results are in agreement with earlier findings that detection of Dynepo™ is best achieved by combining IEF with SDS-PAGE. Haematological parameters were monitored for possible effects due to the long half-life (130 hours) of CERA in blood. Interestingly, although several haematological parameters were significantly changed after the injection of CERA, the endogenous EPO signal was still present in all collected samples. Due to the long half-life and the large size of the CERA molecule (about 60 kDa), it was uncertain whether CERA would be excreted into urine in detectable amounts unless urine collection was preceded by strenuous physical exercise. We find that CERA can be detected in urine without prior exercise in several, but not all, subjects. CERA is nevertheless best detected in serum with regard to both probability and length of detection, in addition to stability in matrix over time.
促红细胞生成素（EPO）因其具有增强运动表现的功效，多年来一直被滥用于运动领域。在过去十年中，通过等电聚焦（IEF）技术，根据内源性和重组 EPO 的不同同工型特征，可以检测到 EPO 的滥用情况。市场上新出现的 EPO，如重组促红细胞生成素 EPO 变体（Dynepo™）和化学修饰的 EPO（CERA），可能会对反兴奋剂斗争带来分析上的挑战。本研究旨在探索检测单次给予 Dynepo™和 CERA 后的可能性和时间窗口。我们的研究结果与早期的研究结果一致，即通过 IEF 与 SDS-PAGE 联合使用可以更好地检测 Dynepo™。监测血液学参数是否因 CERA 在血液中的半衰期（130 小时）较长而受到影响。有趣的是，尽管在注射 CERA 后几个血液学参数发生了显著变化，但所有收集的样本中仍存在内源性 EPO 信号。由于 CERA 的半衰期长且分子较大（约 60 kDa），如果在收集尿液之前没有进行剧烈的体育锻炼，不确定 CERA 是否会以可检测的量排泄到尿液中。我们发现，在没有事先进行剧烈运动的情况下，CERA 可以在一些但不是所有受试者的尿液中被检测到。然而，就检测的可能性和持续时间以及基质稳定性而言，CERA 在血清中的检测效果最好。
