Dou Peng, Liu Zhen, He Jiangang, Xu Jing-Juan, Chen Hong-Yuan
Key Lab of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
J Chromatogr A. 2008 May 9;1190(1-2):372-6. doi: 10.1016/j.chroma.2008.03.001. Epub 2008 Mar 6.
Human erythropoietin (hEPO) is a glycoprotein hormone produced primarily by the kidney, which stimulates red blood cell production. Recombinant human erythropoietin (rhEPO), generally produced in Chinese hamster ovary (CHO) cells, can be used as not only a therapeutic protein but also a doping agent in sports. Profiling of EPO glycoforms is a critical means for quality control in pharmaceutical industrial and anti-doping analysis of misuse in sports. However, the existing methods for the analysis of EPO are associated with either time consuming or poor resolution. In this work, a rapid and high-resolution glycoform profiling method was presented based on capillary isoelectric focusing (cIEF) with whole column imaging detection (WCID). Experimental conditions that influence the separation were investigated. Under optimized conditions, rhEPO from three different sources were resolved into distinct populations within 5 min with excellent reproducibility. As compared with existing methods, the presented method exhibited the advantages of speed and high resolution. If combined with an effective sample enrichment step and a much more sensitive WCID version, the method can be a potential alternative for the detection of rhEPO misuse in sports.
人促红细胞生成素（hEPO）是一种主要由肾脏产生的糖蛋白激素，它能刺激红细胞生成。重组人促红细胞生成素（rhEPO）通常在中国仓鼠卵巢（CHO）细胞中产生，它不仅可以用作治疗性蛋白质，还可以用作体育赛事中的兴奋剂。促红细胞生成素糖型分析是制药工业质量控制以及体育赛事中兴奋剂滥用反兴奋剂分析的关键手段。然而，现有的促红细胞生成素分析方法要么耗时，要么分辨率低。在这项工作中，基于全柱成像检测（WCID）的毛细管等电聚焦（cIEF）技术，提出了一种快速、高分辨率的糖型分析方法。研究了影响分离的实验条件。在优化条件下，来自三种不同来源的rhEPO在5分钟内被分离成不同的群体，重现性良好。与现有方法相比，该方法具有速度快和分辨率高的优点。如果与有效的样品富集步骤以及更灵敏的WCID版本相结合，该方法可能成为检测体育赛事中rhEPO滥用的潜在替代方法。
