达到20万m/z及以上:原生电子捕获 电荷减少 质谱学 解卷 大型生物制药分析物中的异质信号
Kyle I P Le Huray1, Tobias P Wörner2, Tiago Moreira1
1Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Woodhouse Lane, Leeds LS2 9JT, U.K.
ACS central science
|September 2, 2024
概括
电子捕获电荷减小 (ECCR) 通过减少离子电荷状态来增强原生质谱. 这种技术成功地从异质样本 (如病毒囊) 中解复杂的信号,从而使更大的生物分子得到分析.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 质谱测量质量谱测量
背景情况:
- 原生质谱在检测大型生物分子方面表现出色,但由于重叠的信号,与异质样本作斗争.
- 在异质样本中复杂的离子分布对准确的表征提出了重大挑战.
研究的目的:
- 应用电子捕获电荷减值 (ECCR) 来改进大型异质生物分子组件的特性.
- 扩大用于Orbitrap质谱分析的质量与电荷 (m/z) 比率上限.
主要方法:
- 原生ECCR是在细菌的Chaperonin GroEL和腺相关病毒 (AAV) 囊组件上进行的.
- 分析使用Q Exactive UHMR质谱仪进行,仪器进行了轻微修改.
主要成果:
- ECCR显著降低了AAV8体的电荷状态,高达90%,转移信号超过10万m/z.
- 对于高度异质的AAV样本,包括带有遗传载荷的样本,实现了电荷状态分辨率和平均质量确定.
- UHMR仪器检测到超过20万m/z的减电离子,证明了前所未有的m/z上层检测极限.
结论:
- 在原生质谱学中,ECCR是解异质信号的强大工具.
- 这项研究证明了 Orbitrap 仪器记录的最高 m/z 信号,扩大了它对更重的分析物的适用性.
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