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应用新型生物素化克隆 AE7A5 抗体和进一步优化的十二烷基肌氨酸聚丙酰胺凝胶电泳方案进行微量重组人促红细胞生成素给药研究的数据。

Data from a microdosed recombinant human erythropoietin administration study applying the new biotinylated clone AE7A5 antibody and a further optimized sarcosyl polyacrylamide gel electrophoresis protocol.

机构信息

Doping Control Laboratory Seibersdorf, Seibersdorf Labor GmbH, Seibersdorf, Austria.

European Monitoring Center for Emerging Doping Agents, German Sport University Cologne, Cologne, Germany.

出版信息

Drug Test Anal. 2023 Feb;15(2):163-172. doi: 10.1002/dta.2989. Epub 2021 Jan 21.

Abstract

Erythropoietin (EPO) is a hormone, which stimulates the production of red blood cells. Due to its performance-enhancing effect, it is prohibited by the World Anti-Doping Agency (WADA). In order to reduce the detection window of EPO doping, athletes have been applying low doses of recombinant EPO (e.g., <10 IU/kg body weight, daily or every second day) instead of larger doses twice or more per week (e.g., 30 IU/kg). Microdoses of Retacrit (epoetin zeta), an EPO biosimilar, were administered intravenously and subcutaneously to human males and females. Urine and serum samples were collected and analysed applying the new biotinylated clone AE7A5 EPO antibody and a further optimized sarcosyl polyacrylamide gel electrophoresis (SAR-PAGE) protocol. With the improved protocol, microdosed Retacrit (7.5 IU/kg body weight [BW]) was detectable for at least 52 h after intravenous administration. Detection windows were approximately the same for serum and urine and doubled after subcutaneous administration (~104 h). Previous studies applying different electrophoretic techniques and the not further optimized SAR-PAGE protocol revealed considerably shorter detection windows for recombinant human erythropoietin (rhEPO) microdoses. Because the new biotinylated antibody performed significantly more sensitive than the nonbiotinylated version, the new protocol will improve the sensitivity and hence detectability of recombinant EPO in doping control.

摘要

促红细胞生成素(EPO)是一种激素,可刺激红细胞的生成。由于其具有增强表现的作用,因此被世界反兴奋剂机构(WADA)禁止使用。为了减少 EPO 兴奋剂的检测窗口,运动员一直在使用低剂量的重组 EPO(例如,<10IU/kg 体重,每天或每两天一次),而不是更大剂量(例如,每周两次或更多次,30IU/kg)。微剂量的 Retacrit(epoetin zeta),一种 EPO 类似物,被静脉内和皮下施用于男性和女性。收集尿液和血清样本,并应用新的生物素化克隆 AE7A5 EPO 抗体和进一步优化的肌氨酸聚丙酰胺凝胶电泳(SAR-PAGE)方案进行分析。使用改进的方案,静脉内给予的微剂量 Retacrit(7.5IU/kg 体重[BW])可至少在 52 小时后检测到。血清和尿液的检测窗口大致相同,皮下给药后则延长至两倍(~104 小时)。以前的研究应用不同的电泳技术和未进一步优化的 SAR-PAGE 方案,表明重组人促红细胞生成素(rhEPO)微剂量的检测窗口要短得多。由于新的生物素化抗体比非生物素化版本的灵敏度显著更高,因此新方案将提高兴奋剂控制中重组 EPO 的灵敏度和检测能力。

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