Acid/Salt/pH Gradient Improved Resolution and Sensitivity in Proteomics Study Using 2D SCX-RP LC-MS

CORC > 武汉植物园 > 中国科学院武汉植物园

	Acid/Salt/pH Gradient Improved Resolution and Sensitivity in Proteomics Study Using 2D SCX-RP LC-MS
	Wu, JL; Zhu, MZ; Li, N; Wang, YT; Liu, N; Guo, MQ; Sun, BQ; Zhou, H; Liu, L
刊名	JOURNAL OF PROTEOME RESEARCH
	2017
卷号	16 期号:9 页码:3470-3475
英文摘要	The usage of strong cation exchange (SCX) chromatography in proteomics is limited by its poor resolution and nonspecific hydrophobic interactions with peptides, which lead to peptide overlap across fractions and change of peptide retention, respectively. The application of high concentration of salt (up to 1000 mM) in SCX also restricted its use in online 2D SCX-RP LC. In the present research, we first exploited the chromatographic ability of online 2D SCX-RP LC by combination of acid, salt, and pH gradient, three relatively independent modes of eluting peptides from SCX column. 50% ACN was added to elution buffer for eliminating hydrophobic interactions between SCX matrix and peptides, and the concentration of volatile salt was reduced to 50 mM. Acid/salt/pH gradient showed superior resolution and sensitivity as well as uniform distribution across fractions, consequently leading to significant improvements in peptide and protein identification. 112 191 unique peptides and 7373 proteins were identified by acid/salt/pH fractionation, while 69 870 unique peptides and 4536 proteins were identified by salt elution, that is, 62.5 and 60.6% more proteins and unique peptides, respectively, identified by the former. Fraction overlap was also significantly minimized by acid/salt/pH approach. Furthermore, acid/salt/pH elution showed more identification for acidic peptides and hydrophilic peptides.
内容类型	期刊论文
源URL	[http://202.127.146.157/handle/2RYDP1HH/1315]
专题	中国科学院武汉植物园
推荐引用方式 GB/T 7714	Wu, JL,Zhu, MZ,Li, N,et al. Acid/Salt/pH Gradient Improved Resolution and Sensitivity in Proteomics Study Using 2D SCX-RP LC-MS[J]. JOURNAL OF PROTEOME RESEARCH,2017,16(9):3470-3475.
APA	Wu, JL.,Zhu, MZ.,Li, N.,Wang, YT.,Liu, N.,...&Liu, L.(2017).Acid/Salt/pH Gradient Improved Resolution and Sensitivity in Proteomics Study Using 2D SCX-RP LC-MS.JOURNAL OF PROTEOME RESEARCH,16(9),3470-3475.
MLA	Wu, JL,et al."Acid/Salt/pH Gradient Improved Resolution and Sensitivity in Proteomics Study Using 2D SCX-RP LC-MS".JOURNAL OF PROTEOME RESEARCH 16.9(2017):3470-3475.