• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>RSC Advances >Comprehensive quali-quantitative profiling of neutral and sialylated O-glycome by mass spectrometry based on oligosaccharide metabolic engineering and isotopic labeling
【24h】

Comprehensive quali-quantitative profiling of neutral and sialylated O-glycome by mass spectrometry based on oligosaccharide metabolic engineering and isotopic labeling

机译：基于寡糖代谢工程和同位素标记的质谱法对中性和唾液酸化O糖组进行定性和定量分析

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Mass spectrometry (MS) analysis combined with stable isotopic labeling is of great importance for quantitatively profiling abnormal sialylated O -glycans associated with disease development, but technically hindered by the poor releasing efficiency of O -glycans from glycoprotein or the labile nature of sialic acid residues at glycans. Herein, we developed an isotopic precursor based metabolic amplification and labeling (IPMAL) technique for relative quantitative profiling of the repertoire O -glycans between normal and tumor cells by ESI-MS. Two groups of cells were incubated with peracetylated benzyl-α- N -acetylgalactosamine (Ac _(3) GalNAc-α-Bn ~(d0) ) or a heavy labeled peracetylated benzyl-α- N -acetylgalactosamine (Ac _(3) GalNAc-α-Bn ~(d5) ) precursor respectively to amplify the repertoire of O -glycans as Bn ~(d0/d5) - O -glycans which could achieve the quantitative O -glycome analysis by ESI-MS after derivatization. The established method demonstrates desirable feasibility, accuracy (relative error (RE) ≤ 4.20%), reproducibility (coefficient of variation (CV) ≤ 7.61%, n = 3) and good quantitation linearity ( R ~(2) > 0.99, n = 3) for five Bn- O -glycans with 2 orders of magnitude. Finally, the method has been successfully applied to quantitative analysis of the repertoire O -glycome changes between normal human liver cell line L02 and human hepatoma cell line SMMC-7721. Moreover, the α-2,3/2,6 sialic acid isomers of Bn- O -glycans from these two cells have been further quantitatively distinguished when involved a sialic acid specific derivatization procedure.
机译：质谱分析（MS）与稳定的同位素标记相结合对于定量分析与疾病发展相关的异常唾液酸化O聚糖具有非常重要的意义，但是从技术上受到O聚糖从糖蛋白释放效率低或唾液酸残基不稳定的特性的阻碍在聚糖上。本文中，我们开发了一种基于同位素前体的代谢扩增和标记（IPMAL）技术，用于通过ESI-MS对正常细胞与肿瘤细胞之间的全部O-聚糖进行相对定量分析。将两组细胞与过乙酰化的苄基-α-N-乙酰半乳糖胺（Ac _（3）GalNAc-α-Bn〜（d0））或重标记的过乙酰化的苄基-α-N-乙酰基半乳糖胺（Ac _（3）GalNAc -α-Bn〜（d5））前体分别扩增O-聚糖的库，即Bn〜（d0 / d5）-O-聚糖，衍生化后可通过ESI-MS进行定量的O-糖基分析。建立的方法证明了理想的可行性，准确性（相对误差（RE）≤4.20％），重现性（变异系数（CV）≤7.61％，n = 3）和良好的定量线性（R〜（2）> 0.99，n = 3）对于5个Bn-O-聚糖，具有2个数量级。最终，该方法已成功地用于定量分析正常人肝细胞L02和人肝癌细胞SMMC-7721之间O-糖基的变化。此外，当涉及唾液酸特异性衍生化过程时，来自这两个细胞的Bn-O-聚糖的α-2,3/ 2,6唾液酸异构体已被进一步定量区分。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 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