由TMTpro互补离子启用的基于采集的多重复合数据独立蛋白质学
Zicong Wang1, Peng-Kai Liu2, Haiyan Lu1
1School of Pharmacy, University of Wisconsin─Madison, Madison, Wisconsin 53705, United States.
Analytical chemistry
|November 26, 2025
概括
我们开发了一种新的基于3个复合体TMTpro补充离子 (TMTproC) 的DIA策略,用于准确的定量蛋白质组学. 这种方法克服了数据独立获取 (DIA) 中的比率扭曲,并增强了蛋白质覆盖范围.
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:
- 与数据依赖获取 (DDA) 相比,数据独立获取 (DIA) 提供了优越的蛋白质组覆盖和可重复性.
- 在DIA中,多重复合的等标签受到由协同隔离和协同碎片干扰引起的比率扭曲的阻碍.
研究的目的:
- 引入一种基于3个复合体TMTpro互补离子 (TMTproC) 的新型DIA战略.
- 通过利用互补的离子,在DIA中实现准确的量化,而不会增加光谱复杂性.
主要方法:
- 实施了TMTproC标签,在互补离子之间设置了4-Da间距,以最大限度地减少光谱重叠.
- 优化了高能碰撞解离 (HCD) 设置,用于互补的离子生成.
- 验证了使用三性牛血清白蛋白 (BSA) 和酵母蛋白质样本的方法.
主要成果:
- 实现了BSA的中位数水平比值在预期值的10%以内.
- 在三张复制品中,证明的变化系数 (CVs) 中位数低于4%.
- 在酵母蛋白质组分析中,在10倍的动态范围中展示了高量化精度和最小的比率扭曲.
结论:
- TMTproC-DIA是一种强大的多功能策略,用于准确的定量蛋白质组学.
- 这种方法有效地减少了比率扭曲,并增强了DIA中的蛋白质覆盖范围.
- TMTproC-DIA方法对于高通量蛋白质组学研究是可扩展的.
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