• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>other >Relative Quantitation of Neuropeptides Over a Thousand-fold Concentration Range
【2h】

Relative Quantitation of Neuropeptides Over a Thousand-fold Concentration Range

机译：神经肽逾千倍浓度范围相对定量

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Neuropeptides are essential cell-to-cell signaling molecules that influence diverse regulatory and behavioral functions within biological systems. Differing in their amino acid sequences and posttranslational modifications, hundreds of neuropeptides are produced via a series of enzymatic processing steps, and their levels vary with location, time, and physiological condition. Due to their wide range of endogenous concentrations and inherent chemical complexity, using mass spectrometry (MS) to accurately quantify changes in peptide levels can be challenging. Here we evaluate three different MS systems for their ability to accurately measure neuropeptide levels: capillary liquid chromatography-electrospray ionization-ion trap (CapLC-ESI-IT) MS, ultraperformance liquid chromatography- electrospray ionization-quadrupole-time-of-flight (UPLC-LC-ESI-Q-TOF) MS, and matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) MS. Specifically, eight sample mixtures composed of five neuropeptide standards, with four technical replicates of each, were labeled with H4/D4-succinic anhydride, followed by relative peptide quantitation using the three MS platforms. For these samples, the CapLC-ESI-IT MS platform offered the most robust ability to accurately quantify peptides over a concentration range of 1200-fold, although it required larger sample sizes than the other two platforms. Both the UPLC-ESI-Q-TOF MS and the MALDI-TOF MS systems had lower limits of quantification, with the MALDI-TOF having the lowest. By implementing several data acquisition schemes and optimizing the data analysis approaches, we were able to accurately quantify peptides over a three orders of magnitude concentration range using either the UPLC or MALDI-TOF platforms. Overall these results increase our understanding of both the capabilities and limits of using MS-based approaches to measure peptides.
机译：神经肽是必不可少的细胞间信号分子，会影响生物系统内各种调节和行为功能。氨基酸序列和翻译后修饰不同，通过一系列酶促加工步骤可产生数百种神经肽，并且它们的水平会随位置，时间和生理状况而变化。由于它们的内源性浓度范围广且固有的化学复杂性，因此使用质谱（MS）准确定量肽水平的变化可能具有挑战性。在这里，我们评估了三种不同的质谱系统准确测量神经肽水平的能力：毛细管液相色谱-电喷雾电离离子阱（CapLC-ESI-IT）质谱，超高效液相色谱-电喷雾电离-四极杆飞行时间（UPLC） -LC-ESI-Q-TOF）MS和基质辅助激光解吸/电离飞行时间（MALDI-TOF）MS。具体来说，用H4 / D4-丁二酸酐标记由五个神经肽标准品组成的八种样品混合物，每种均具有四个技术重复，然后使用三个MS平台进行相对肽定量。对于这些样品，尽管CapLC-ESI-IT MS平台需要比其他两个平台更大的样本量，但它提供了最强大的功能，可以在1200倍的浓度范围内准确定量多肽。 UPLC-ESI-Q-TOF MS系统和MALDI-TOF MS系统均具有较低的定量限，而MALDI-TOF的定量限最低。通过实施几种数据采集方案并优化数据分析方法，我们能够使用UPLC或MALDI-TOF平台在三个数量级浓度范围内准确定量肽。总体而言，这些结果使我们对使用基于MS的方法测量肽的能力和局限性有了更深入的了解。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

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