Targeted quantitation of proteins by mass spectrometry.

Targeted quantitation of proteins by mass spectrometry.
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DOI:
10.1021/bi400110b
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发表时间:
2013-06-04
期刊:
影响因子:
2.9
通讯作者:
Zimmerman LJ
Zimmerman LJ
中科院分区:
生物学3区
文献类型:
--
作者:
Liebler DC;Zimmerman LJ

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蛋白质的定量测量是生物化学实验室中最基本的分析任务之一,但广泛使用的免疫化学方法通常具有有限的特异性和高测量变化。在这篇综述中,我们讨论了应用多反应监测(MRM)质谱,它允许灵敏,精确的定量分析肽和蛋白质,它们来自。肽质谱和序列的数据库、用于分析设计和数据分析的软件工具以及串联质谱仪技术的快速发展允许MRM测定的系统开发。MRM检测的主要优势是靶向特定肽序列(包括变体和修饰形式)的能力,以及允许分析数十至数百种肽的多路复用能力。不同的定量标准化方法提供了平衡精度、灵敏度和测定成本的选择。通过MRM和相关质谱方法进行的靶向蛋白质定量可以通过将方法转化为蛋白质测量来推进生物化学。
Quantitative measurement of proteins is one of the most fundamental analytical tasks in a biochemistry laboratory, but widely used immunochemical methods often have limited specificity and high measurement variation. In this review, we discuss applications of multiple-reaction monitoring (MRM) mass spectrometry, which allows sensitive, precise quantitative analyses of peptides and the proteins from which they are derived. Systematic development of MRM assays is permitted by databases of peptide mass spectra and sequences, software tools for analysis design and data analysis, and rapid evolution of tandem mass spectrometer technology. Key advantages of MRM assays are the ability to target specific peptide sequences, including variants and modified forms, and the capacity for multiplexing that allows analysis of dozens to hundreds of peptides. Different quantitative standardization methods provide options that balance precision, sensitivity, and assay cost. Targeted protein quantitation by MRM and related mass spectrometry methods can advance biochemistry by transforming approaches to protein measurement.
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