通过免疫亲和液体染色学――三重四极质谱学提高蛋白质定量化的灵敏度,使用代过渡总和技术
Jay S Johnson1, Joe Palandra1, Nikolaos Psychogios1
1Pharmacokinetics, Dynamics & Metabolism (PDM), Pfizer, Andover, Massachusetts 01810, United States.
Analytical chemistry
|August 26, 2024
概括
回声总和显著提高了使用免疫亲和丰富和纳米液体染色学-双重质谱法 (IA-nLC-MS/MS) 的蛋白质量测量灵敏度. 这种技术提高了人类血清中低丰度蛋白质标的准确性和精度.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 质谱测量质量谱测量
背景情况:
- 液体染色学-并联质谱学 (LC-MS/MS) 的进展旨在降低低丰度蛋白质的量化极限.
- 免疫亲和 (IA) 丰富和纳米流LC (nLC) 是已建立的方法来提高灵敏度.
- 对于敏感蛋白质分析,信号与噪声 (S/N) 比率的进一步改进通常是必要的.
研究的目的:
- 为了评估回声总和对蛋白质量定量的适用性,使用IA与nLC-MS/MS (IA-nLC-MS/MS) 结合.
- 评估回声总结对人类血清中蛋白质测定准确度,精度和灵敏度的影响.
主要方法:
- 应用回声总和,是一种涉及重复收集和总和单一选择反应监测 (SRM) 过渡的技术.
- 使用了连续的IA丰富与nLC-MS/MS结合用于蛋白质分析.
- 在人血清样本中量化β神经生长因子 (NGF) 和可溶性亚亚糖蛋白受体 (sASGPR).
主要成果:
- 与传统方法相比,回声总和的五次代提高了NGF量化准确度和精度.
- 反响总结大约使sASGPR的功能灵敏度翻了一番.
- 通过回声总和,sASGPR的最小可量化的水平被扩展了4倍.
结论:
- 反响总和在IA-nLC-MS/MS测定中直接适用于蛋白质定量,并且对蛋白质定量有好处.
- 该技术为低丰度蛋白质标提供了显著的灵敏度,准确度和精度改进.
- 对于大多数蛋白质IA-nLC-MS/MS应用,预计会有类似的灵敏度增长.
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