Characterization and Detection of Erythropoietin Fc Fusion Proteins Using Liquid Chromatography–Mass Spectrometry,Journal of Proteome Research

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Characterization and Detection of Erythropoietin Fc Fusion Proteins Using Liquid Chromatography–Mass Spectrometry
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2017-12-19 00:00:00 , DOI: 10.1021/acs.jproteome.7b00739
Natalia V. Mesonzhnik ₁ , Pavel V. Postnikov ₂ , Svetlana A. Appolonova ₁ , Grigory I. Krotov ₃

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Erythropoietin Fc (EPO-Fc) fusion proteins are potential drug candidates that have been designed for the treatment of anemia in humans by stimulating erythrocyte production. Such compounds can be considered performance-enhancing agents that may be used by athletes in endurance sports. This study describes the primary structure of commercially available EPO-Fc based on comprehensive liquid chromatography coupled with mass spectrometry (LC–MS) analysis. A bottom-up approach and the intact molecular weight (MW) measurement of deglycosylated protein and its IdeS proteolytic fractions was used to determine the amino acid sequence of EPO-Fc. Using multiple proteases, peptides covering unknown fusion breakpoints (spacer peptides) were identified. We demonstrated that “spacer peptides” can be used in the determination of EPO-Fc fusion proteins in biological samples using common LC–tandem MS methods.

中文翻译：

液相色谱-质谱法表征和检测促红细胞生成素Fc融合蛋白

促红细胞生成素Fc（EPO-Fc）融合蛋白是潜在的候选药物，已被设计用于通过刺激红血球生成来治疗人类贫血。这样的化合物可以被认为是运动员在耐力运动中可以使用的性能增强剂。这项研究描述了基于全面液相色谱和质谱分析（LC-MS）的市售EPO-Fc的主要结构。使用自下而上的方法和去糖基化蛋白及其IdeS蛋白水解级分的完整分子量（MW）测量来确定EPO-Fc的氨基酸序列。使用多种蛋白酶，鉴定了覆盖未知融合断裂点的肽（间隔肽）。

更新日期：2017-12-19

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