• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of mass spectrometry: JMS >Low LC-MS/MS detection of glycopeptides released from pmol levels of recombinant erythropoietin using nanoflow HPLC-chip electrospray ionization
【24h】

Low LC-MS/MS detection of glycopeptides released from pmol levels of recombinant erythropoietin using nanoflow HPLC-chip electrospray ionization

机译：使用纳流HPLC芯片电喷雾电离低LC-MS / MS检测从重组人促红细胞生成素的pmol水平释放的糖肽

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The test used by anti-doping laboratories to detect the misuse of recombinant erythropoietin (rhEPO) is based on its different migration pattern on isoelectric focusing (IEF) gel compared with the endogenous human erythropoietin (hEPO) that can possibly be explained by structural differences. While there is definitely a need to identify those differences by LC-MS/MS, the extensive characterization that was achieved for the rhEPO was never performed on human endogenous EPO because its standard is not available in sufficient amount. The goal of this study was to develop an analytical method to detect pmol amounts of N-linked and O-linked glycopeptides of the recombinant hormone as a model. Using a nanoflow HPLC-Chip electrospray ionization/ion trap mass spectrometer, the diagnostic ion at m/z 366 of oligosaccharides was monitored in the product ion spectra to identify the four theoretical glycosylation sites, Asn24, Asn38, Asn83 and Ser126, respectively, on glycopeptides 22-37, 38-55, 73-96 and 118-136. With 3 pmol of starting material applied on Chip, only the desialylated N-glycopeptides 22-37 and 38-55/38-43 could be observed, and of all the glycan isoforms, those with the smaller structures were predominantly detected. While the preservation of the sialic acid moieties decreased the detection of all the N-glycopeptides, it allowed a more extensive characterization of the O-linked glycopeptide 118-136. The technique described herein provides a mean to detect glycopeptides from commercially available pharmaceutical preparations of rhEPO with the sensitivity required to analyze pmol amounts of hEPO, which could ultimately lead to the identification of structural differences between the recombinant and the human forms of the hormone.
机译：反兴奋剂实验室用于检测重组促红细胞生成素（rhEPO）滥用的测试是基于其在等电聚焦（IEF）凝胶上与内源性人类促红细胞生成素（hEPO）相比的不同迁移模式，这可能可以通过结构差异来解释。虽然绝对有必要通过LC-MS / MS识别这些差异，但对于rhEPO所实现的广泛表征从未在人类内源性EPO上进行过，因为其标准品数量不足。这项研究的目的是开发一种分析方法，以检测pmol量的重组激素作为模型的N-联和O-联糖肽。使用纳流HPLC-Chip电喷雾电离/离子阱质谱仪，在产物离子光谱中监测m / z 366寡糖的诊断离子，以分别鉴定出四个理论糖基化位点Asn24，Asn38，Asn83和Ser126。糖肽22-37、38-55、73-96和118-136。在芯片上施加3 pmol起始原料后，只能观察到脱唾液酸化的N-糖肽22-37和38-55 / 38-43，并且在所有聚糖同工型中，主要检测到结构较小的那些。虽然唾液酸部分的保存减少了所有N-糖肽的检测，但它允许更广泛地表征O-连接的糖肽118-136。本文所述的技术提供了一种手段，用于以可分析pmol量的hEPO所需的灵敏度检测来自rhEPO的市售药物制剂中的糖肽，这最终可能导致鉴定重组形式和人形式的激素之间的结构差异。

著录项

相似文献

  • 外文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号