蛋白质组学最佳碰撞能量:离子移动性分离的影响
Kinga Nagy1,2, Gabriella Gellén3, Dávid Papp2,3
1MS Proteomics Research Group, Research Centre for Natural Sciences, Budapest, H-1117, Hungary.
Journal of mass spectrometry : JMS
|July 7, 2023
概括
降低碰撞能量 (CE) 在使用离子流动性光谱法 (IMS) 与LC-MS/MS相结合时可以提高鉴定得分. 优化CE可以通过IMS分离来增强自下而上的蛋白质组学分析.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 分析化学 分析化学
- 分离科学 分离科学
背景情况:
- 离子移动谱法 (IMS) 是一种分离技术,通常与LC-MS/MS相结合.
- 由于气体碰撞,IMS可能会导致离子加热,这可能会影响质谱分析.
- 了解IMS中的离子行为对于优化自下而上的蛋白质组学至关重要.
研究的目的:
- 为了研究离子移动性光谱对碰撞能量 (CE) 优化对自下而上的蛋白质组学的影响.
- 为了确定最佳的CE设置,用于用IMS和没有IMS的鉴定.
- 为建立带连字符的IMS质谱平台提供实用指导.
主要方法:
- 在循环离子流动性质谱仪上对来自HeLa消化物的三性的LC-MS/MS分析.
- 碰撞能量的系统变化 (CE) 有或没有IMS.
- 使用Byonic搜索引擎进行体识别得分分析.
主要成果:
- 当IMS应用时,较低的CE值 (平均6.3V减少) 显著改善了鉴定得分.
- 最佳的CE设置在具有IMS和没有IMS的设置之间有所不同.
- 当使用IMS时,制造商建议的CE参数通常过高.
结论:
- 优化碰撞能量对于最大限度地提高IMS合LC-MS/MS中的类鉴定至关重要.
- 与标准LC-MS/MS相比,IMS分离需要较低的CE设置.
- 这些发现有助于实际实施和优化IMS增强的蛋白质组学工作流程.
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