合高场不对称波形离子移动性光谱与毛细管区电泳-双重质谱用于上下蛋白质学
Tian Xu1, Qianjie Wang1, Qianyi Wang1
1Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, Michigan 48824, United States.
Analytical chemistry
|May 31, 2023
概括
高场不对称波形离子流动性光谱学 (FAIMS) 与毛细管区电泳-双重质谱学 (CZE-MS/MS) 相结合,显著提高了自上而下的蛋白质组学中的蛋白形识别. 这种先进的技术提高了对较大的蛋白质形式的检测,并提高了整体灵敏度.
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 生化学
背景情况:
- 毛细管区电泳-双重质谱 (CZE-MS/MS) 对于自上而下的蛋白质组学 (TDP) 是至关重要的.
- 目前的TDP方法在分离和检测各种蛋白质形式,特别是较大的蛋白质形式方面面临挑战.
研究的目的:
- 通过整合在线气相蛋白形分离来提高TDP的CZE-MS/MS性能.
- 评估高场不对称波形离子流动性光谱法 (FAIMS) 在蛋白形分离和识别方面的有效性.
主要方法:
- 将CZE-MS/MS与FAIMS结合起来,用于在线蛋白形分离.
- 在FAIMS中使用不同的补偿电压 (CV) 来根据尺寸分成蛋白质形状.
- 分析酵母样本,以比较CZE-FAIMS-MS/MS与单独的CZE-MS/MS.
主要成果:
- FAIMS 分割将已识别的蛋白质形式的中位质量从<10 kDa转移到~30 kDa,表明基于大小的分离.
- 与单独使用CZE-MS/MS相比,CZE-FAIMS-MS/MS增加了近3倍的蛋白型鉴定.
- 在20-45kDa范围内的蛋白形体的识别得到了6倍的改进,信号噪声比增加了近20倍.
结论:
- CZE-FAIMS-MS/MS有效地按大小分化蛋白质形式,显著提高TDP的灵敏度和覆盖范围.
- 这种整合代表了毛细管电泳与用于TDP的离子移动性谱学的首次合.
- 开发的CZE-FAIMS-MS/MS技术是推动蛋白形状分析的宝贵工具.
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