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相关概念视频

Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

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Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
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双步溶解增加了高灵敏度纳米HILIC/MS/MS基础蛋白质的覆盖范围.

Koshin Akamatsu1, Eisuke Kanao1,2, Ayana Tomioka1

  • 1Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.

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概括

我们开发了一种两步溶解方法,以改进纳米级的水友互动色谱学,并配合双重质谱学 (nanoHILIC/MS/MS). 这种技术增强了的溶解性,并促进了蛋白质组学研究中的蛋白质识别.

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科学领域:

  • 蛋白质组学是指蛋白质组学.
  • 分析化学 分析化学
  • 生物化学 生物化学

背景情况:

  • 纳米级的水友互动色谱学与双重质谱学 (纳米HILIC/MS/MS) 结合,为蛋白质学提供了潜力.
  • 在有机溶剂丰富的溶液中的溶解性是纳米HILIC/MS/MS的主要限制.
  • 反相液态色谱 (RPLC) 是一种常见的,但有时不那么敏感的方法.

研究的目的:

  • 为纳米HILIC/MS/MS.开发一种改进的化溶解方法.
  • 为了提高的溶解性和与高度有机溶剂的兼容性.
  • 为了提高纳米HILIC/MS/MS的灵敏度和范围,用于蛋白质组学应用.

主要方法:

  • 开发了一种两步溶解程序:初始溶解在25%的乙二 (ACN),然后稀释到95%的ACN.
  • 用纳米HILIC/MS/MS分析了类样本,并与95%ACN的直接溶解相比较.
  • 性能与纳米尺度逆相液态染色学结合使用HeLa细胞三性的双重质谱法 (nanoRPLC/MS/MS) 进行了基准测试.

主要成果:

  • 两步方法在纳米HILIC/MS/MS中显著提高了的溶解度和强度,82.8%的显示强度增加 (平均增益为20.9%).
  • 与使用最小样本 (2.5 ng) 的纳米RPLC/MS/MS相比,使用 nanoHILIC/MS/MS 的新方法确定了8.47倍多的和3.54倍多的蛋白质组.
  • 在ACN丰富的移动相中,增强的载荷和优异的电喷离电离 (ESI) 效率有助于高灵敏度.

结论:

  • 开发的两步溶解方法有效地克服了纳米HILIC/MS/MS中的类溶解度限制.
  • 这种方法显著提高了灵敏度,使得蛋白质覆盖范围更深.
  • 优化的nanoHILIC/MS/MS系统为敏感蛋白质组学提供了一个强大的平台,包括临床和单细胞分析.