Quantification Analysis of Targeted Proteins in Complex Sample by Novel Data Independent Acquisition
10.11895/j.issn.0253-3820.140365
- VernacularTitle:创新数据非依赖性采集用于复杂基质目标蛋白质的定量分析
- Author:
Wei ZHANG
;
Kiyonami REIKO
;
Zheng JIANG
;
Wei CHEN
- Publication Type:Journal Article
- Keywords:
Orbitrap;
Data independent acquisition;
Proteomics;
Absolute quantification
- From:Chinese Journal of Analytical Chemistry
2014;(12):1750-1758
- CountryChina
- Language:Chinese
-
Abstract:
Data independent acquisition ( DIA ) is a novel MS scan mode for quantitative proteomics, acquiring all precursors as well as fragments without any loss of low abundant ions, and breaks the throughput limitation of product ion quantification by high-resolution MS. Here we developed three DIA methods on quadrupole-linear ion trap-Orbitrap ( Q-qIT-OT ) Tribrid MS, classic DIA, as well as novel wide isolation window SIM scan ( WiSIM)-DIA and full scan-DIA ( Full MS-DIA) . Quantitative analysis of 10 low abundant peptides spiked in Hela cell digest was performed by the three methods for linearity, reproducibility and sensitivity evaluation. The results showed that the LOQs reached amol level ( 14-435 amol ) with good linearity and effective MS/MS confirmation. WiSIM-DIA utilizes ultra-high resolution SIM scan for quantification complementary with classic DIA. The isolation window of Full MS-DIA was down to 3 amu, and the data could be directly used for database searching, thus realizing the integration of data dependent acquisition ( DDA) and DIA, and avoiding the limitation of using spectra library.
