纳米电子喷射电离发射器位置对非故意源内激活和蛋白质离子的影响
Samantha O Shepherd1, Austin W Green1, Elizabeth S Resendiz1
1Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon 97403-1253, United States.
Journal of the American Society for Mass Spectrometry
|February 20, 2024
概括
在原生离子移动性质谱学 (IM-MS) 中的发射器位置显著影响碰撞诱导的展开/解离 (CIU/D) 实验. 微小的调整可以改变8V的能量潜力,影响蛋白质结构分析.
科学领域:
- 分析化学 分析化学
- 生物物理化学 生物物理化学
- 质谱测量质量谱测量
背景情况:
- 原生离子移动性质谱法 (IM-MS) 使用纳米电子喷雾电离 (nESI) 将蛋白质离子引入气相.
- nESI设置通常包括用于信号调和和控制 adduct 形成的移动阶段.
- 发射器位置调整虽然对信号优化有用,但可能无意中影响下游CIU/D实验.
研究的目的:
- 调查纳米电子喷雾电离 (nESI) 发射器位置对原生离子移动性质谱 (IM-MS) 实验中的碰撞诱导的展开/解离 (CIU/D) 发射器位置的影响.
- 量化发射器位置对分解曲线和结构过渡的中点潜力 (CID50/CIU50) 的影响.
主要方法:
- 在商用质谱仪上,nESI发射器位置的系统变化 (Waters Synapt G2-Si,Agilent 6545XT).
- 对各种分析物 (全体球蛋白,白蛋白脑内素,Shiga毒素1亚单元B,牛血清白蛋白,NIST mAb) 的碰撞诱导解离 (CID) 和碰撞诱导展开 (CIU) 分解曲线的分析.
- 与发射器位置相对应的CID50转移的映射.
- 使用CIU指纹和CIU50转移的RMSD对源内激活的评估.
主要成果:
- 在nESI发射器位置的小变化可以将CID50/CIU50潜力转移多达8V.
- 离仪器进口更近的发射器位置通常会导致Waters Synapt G2-Si.的源内激活更大.
- 观察到的效应在仪器模型 (Waters vs. Agilent) 和分析品之间有所不同.
- 通过分析CIU指纹和结构转变潜力,确认了源内激活.
结论:
- 精确定位的nESI发射器是一个关键参数,显著影响CIU/D实验在IM-MS.
- 发射器位置的不计数变化可能导致无法重现的结果和对蛋白质结构动态的误解.
- 精心优化和记录发射器位置对于可靠和可比的CIU/D数据采集至关重要.
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