文献类型：期刊文献

英文题名：Comparative Evaluation of Small Molecular Additives and Their Effects on Peptide/Protein Identification

作者：Gao, Jing[1,2,3];Zhong, Shaoyun[1,2,3];Zhou, Yanting[2,3,5];He, Han[2,3];Peng, Shuying[6];Zhu, Zhenyun[2,3];Liu, Xing[2,3];Zheng, Jing[5];Xu, Bin[1];Zhou, Hu[2,3,4,7]

机构：[1]Shanghai Univ, Coll Sci, Dept Chem, Shanghai 200444, Peoples R China;[2]Chinese Acad Sci, Dept Analyt Chem, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China;[3]Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China;[4]Univ Chinese Acad Sci, Beijing 100049, Peoples R China;[5]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, 130 Meilong Rd, Shanghai 200237, Peoples R China;[6]Thermo Fisher Sci China Co Ltd, 6 Bldg,27 Xinjinqiao Rd, Shanghai 201206, Peoples R China;[7]Shanghai Univ Tradit Chinese Med, E Inst Shanghai Municipal Educ Comm, 1200 Cai Lun Rd, Shanghai 201203, Peoples R China

年份：2017

卷号：89

期号：11

起止页码：5785

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20172503808262);WOS:【SCI-EXPANDED(收录号:WOS:000403036800023)】；

基金：This research was supported by Grants 21375138 and 31570830 from the National Natural Science Foundation of China; grant 2013ZX09507001 from the Ministry of Science and Technology of China; the "one hundred talent program" of the Chinese Academy of Sciences; the Strategic Priority Research Program (Grant XDA12030203) of the Chinese Academy of Sciences; the Innovation Project of Instrument and Equipment Function Development (Grant YZ201542) and the Innovation Project of Instrument and Equipment Function Development of the Chinese Academy of Sciences (Development of a background noise reduction device for nano-electrospray ion source of mass spectrometry) from the Bureau of Goods, Chinese Academy of Sciences; E-institute of Shanghai Municipal Education Committee E03008; and Grant JKLPRD201404 from the Open Project Program of Jiangsu Key Laboratory of Pediatric Respiratory Disease, Nanjing University of Chinese Medicine.

语种：英文

外文关键词：Mass spectrometry - Quality control - Peptides - Acetone - Soaps (detergents) - Urea

摘要：Detergents and salts are widely used in lysis buffers to enhance protein extraction from biological. samples, facilitating in-depth proteomic analysis., However, these detergents and Salt additives must be efficiently removed from the digested samples prior to LC-MS/MS analysis to obtain high-quality mass spectra. Although filter-aided sample preparation (FASP), acetone precipitation (AP), followed by in-solution digestion,, and strong cation exchange-based centrifugal proteomic reactors (CPRS) are commonly used for proteomic sample processing, little is-known about their efficiencies at,removing detergents and salt additives. In this study, an integrative workflow for the quantification of small molecular additives in proteomic. sample's, developing a multiple reaction monitoring (MRM)-based LC-MS approach for the quantification of six additives (i.e., Tris, urea, CHAPS, SDS, SDC; and Triton X-100) and (ii) systematically evaluated the relationships between the level of additive remaining: in samples following sample processing and the number of peptides/proteins identified by mass Spectrometry. Although FASP outperformed the-other two methods, the results were complementary in terms of peptide/protein identification, as well as the GRAVY index and amino acid distributions. This is the first systematic and quantitative Study-of the effect of detergents and salt, additives on protein identification. This MRM-based approach can be used for an unbiased evaltation,of the performance of new sample preparation Method's. Data are available via PotegmeXchange under identifier PXD005405.