Ion-pair Reversed-phase×Low-pH Reversed-phase Two-dimensional Liquid Chromatography for In-depth Proteomic Profiling  

作　　者：NIU Wenxue LIU Zheyi LIU Jing LAI Can ZHANG Tingting ZHAO Heng WANG Guosheng WANG Fangjun 

机构地区：[1]College of Chemical Engineering,Shenyang University of Chemical Technology,Shenyang,110142,P.R.China [2]CAS Key Laboratory of Separation Sciences for Analytical Chemistry,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian,116023,P.R.China [3]College of Pharmacy,Dalian Medical University,Dalian,116044,P.R.China [4]University of Chinese Academy of Sciences,Beijing,100049,P.R.China

出　　处：《Chemical Research in Chinese Universities》2023年第2期260-265,共6页高等学校化学研究（英文版）

基　　金：National Key R&D Program of China(No.2019YFE0119300);National Natural Science Foundation of China(No.32088101);Original Innovation Project of CAS(No.ZDBS-LY-SLH032);Project of Dalian Institute of Chemical Physics,CAS,China(No.DICPI202007).

摘　　要：High-resolution liquid chromatography separation is essential to in-depth proteomic profiling of complex biological samples.Herein,we established an ion-pair reversed-phase×reversed-phase two-dimensional liquid chromatography(IPRP×RP 2DLC)strategy for comprehensive proteomic analysis.Both RPLC separation dimensions were performed at low pH,with trifluoroacetic acid(TFA)and formic acid(FA)as mobile phase addictive,respectively.As the good separation resolution offered by ion-pairing effect of TFA,the fractionation efficiency was greatly improved with 74.0%peptides identified in just one fraction.Comparing with conventional high pH RP fractionation,the overall separation rate of IPRP was about 1.6 times that of high-pH RP,which increased the number of identified peptides by 21%.Further,2169 proteins and 8540 peptides were confidently identified from crude serum sample by our IPRP×RP 2DLC strategy,exhibiting great potential in clinical proteomics in the future.

关 键 词：Two-dimensional liquid chromatography(2DLC) Ion-pair reversed-phase liquid chromatography(IPRP-LC) IPRP×low-pH RP 2DLC PROTEOMICS Mass spectrometry 

分 类 号：O652.63[理学—分析化学]