Rapid and high-resolution glycoform profiling of recombinant human erythropoietin by capillary isoelectric focusing with whole column imaging detection
Rapid and high-resolution glycoform profiling of recombinant human erythropoietin by capillary isoelectric focusing with whole column imaging detection
复制标题
通过毛细管等电聚焦和全柱成像检测对重组人促红细胞生成素进行快速、高分辨率的糖型分析
DOI：
10.1016/j.chroma.2008.03.001
复制
发表时间：
2008-05-09
影响因子：
4.1
通讯作者：
Chen, Hong-Yuan
中科院分区：
文献类型：
--
作者：
Dou, Peng;Liu, Zhen;Chen, Hong-Yuan
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，它刺激红细胞的产生。重组人促红细胞生成素（rhEPO）一般产生于中国仓鼠卵巢（CHO）细胞中，它不仅可以作为一种治疗蛋白，还可以作为一种体育兴奋剂。EPO糖型分析是医药工业质量控制和体育运动中兴奋剂滥用反分析的重要手段。然而，现有的EPO分析方法要么耗时，要么分辨率差。本文提出了一种基于毛细管等电聚焦（cIEF）和全柱成像检测（WCID）的快速、高分辨率糖型分析方法。研究了影响分离的实验条件。在优化条件下，3种不同来源的rhEPO在5 min内分离成不同的群体，重现性良好。与现有方法相比，该方法具有速度快、分辨率高的优点。如果结合有效的样品富集步骤和更敏感的WCID版本，该方法可以成为检测运动中rhEPO滥用的潜在替代方法。(C) 2008 Elsevier B.V.版权所有
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. (C) 2008 Elsevier B.V. All rights reserved.