Erythropoietin (EPO) has protective effects in several tissues and could be used for therapeutic purposes, but the doses of EPO that can be beneficial in case of hypoxic–ischemic conditions due to overinduced erythropoiesis could be detrimental in treated patients. Carbamylation of erythropoietin maintains the tissue‐protective effects of EPO but without erythropoietic effects. Carbamylated EPO (CEPO) is listed in WADA Prohibited List in class S2 as “Innate repair receptor agonists.” The CEPO was synthesized using the method described previously. Digestion with endoproteinase Lys‐C was used to distinguish rhEPO from CEPO. The digested samples containing recombinant EPO, urinary EPO (uEPO), or CEPO were analyzed by the SAR‐PAGE method (sarcosyl polyacrylamide gel electrophoresis‐PAGE). Endoproteinase Lys‐C breaks the peptide chains of lysine. Lysine residues, converted to homocitrulline by carbamylation, cannot be cleaved by endoproteinase Lys‐C. Therefore, the CEPO protein chain remained unchanged in contrast to rhEPO and uEPO, which allows for easily differentiation of them.

中文翻译：

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用于兴奋剂控制目的的氨基甲酰化促红细胞生成素检测分析方法

促红细胞生成素 (EPO) 在多种组织中具有保护作用，可用于治疗目的，但在过度诱导红细胞生成导致缺氧缺血情况下有益的 EPO 剂量可能对治疗患者有害。促红细胞生成素的氨基甲酰化保持了 EPO 的组织保护作用，但没有促红细胞生成作用。氨基甲酰化 EPO (CEPO) 作为“先天修复受体激动剂”被列入 WADA 禁用清单中的 S2 类。CEPO ​​是使用先前描述的方法合成的。使用内切蛋白酶 Lys-C 进行消化以区分 rhEPO 和 CEPO。通过 SAR-PAGE 方法（肌基聚丙烯酰胺凝胶电泳-PAGE）分析含有重组 EPO、尿 EPO (uEPO) 或 CEPO ​​的消化样品。内切蛋白酶 Lys-C 破坏赖氨酸的肽链。通过氨甲酰化转化为高瓜氨酸的赖氨酸残基不能被内切蛋白酶 Lys-C 切割。因此，与rhEP​​O 和uEPO 相比，CEPO ​​蛋白链保持不变，这使得它们易于区分。