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使用野外不对称离子移动性光谱学改进完整的蛋白质形式的无标签量化

Jake T Kline¹, Michael W Belford², Jingjing Huang²

¹Department of Biology, University of Oklahoma, Norman, Oklahoma 73019, United States.

Analytical chemistry

|May 30, 2023

概括

使用野外不对称离子流动性光谱法 (FAIMS) 的气相分化显著提高了完整蛋白质形式的识别和量化. 这种先进的技术可以改善无标签蛋白质组分析,而不会影响准确度.

科学领域:

蛋白质组学是指蛋白质组学.
分析化学分析化学
生物化学生物化学

背景情况:

完整的蛋白质形式的高通量定量对于理解细胞过程至关重要.
无标签方法很常见,但受样本复杂性的限制,特别是0-30kDa范围内的蛋白质.
现有的分离技术,如HPLC和CE,难以解决复杂的蛋白形混合物.

研究的目的:

用气相分化 (GPF) 来对大肠杆菌的蛋白质形式进行无标签量化的基准.
评估场非对称离子移动性光谱法 (FAIMS) 对蛋白质组分析的影响.
评估GPF是否可以在不牺牲准确性的情况下改善蛋白质形状的识别和量化.

主要方法:

应用气相分化 (GPF) 通过野外不对称离子移动性谱法 (FAIMS) 对大肠杆菌蛋白质形式.
利用最近在Orbitrap仪器仪表方面的进展,以获得高质量的完整和碎片化质谱.
将多个FAIMS补偿电压集成到一个单一的液体染色学-并联质谱实验中.

主要成果:

通过FAIMS的GPF大大增加了已识别的蛋白质形式的数量.
量化蛋白质形式的数量也显著增加.
与没有GPF的传统无标签方法相比,量化准确度保持不变.

结论:

基于FAIMS的GPF是一种有效的策略,可以在无标签的蛋白质量定量中克服样本的复杂性.
这种方法扩大了可以识别和量化的蛋白质形式的范围.
通过将FAIMS与先进仪器集成,可以提高蛋白质组分析能力.

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