期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1216, 期 8, 页码 1270-1278出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2008.10.002
关键词
Peptidomics; Ordered porous nanoparticles; Strong ion exchange; Reversed phase; Two-dimensional separation; Mouse liver
资金
- National Natural Science Foundation of China [20735004, 20675081, 20675038]
- State Key Basic Research Program [2005CB522701, 2007CB914102]
- High Technology Research Program [2006AA02A309]
- Ministry of Science and Technology of China [KJCX2.YW.H09]
- Chinese Academy of Sciences, and the Knowledge Innovation Program of Dalian Institute of Chemical Physics to H. Zou
We report the development of a combined strategy for high capacity, comprehensive enrichment of endogenous peptide from complex biological samples at natural pH condition. MCM-41 nanoparticles with highly ordered nanoscale pores (i.e. 4.8 nm) and high-surface area (i.e. 751 m(2)/g) were synthesized and modified with strong cation-exchange (SCX-MCM-41) and strong anion-exchange (SAX-MCM-41) groups. The modified nanoparticles demonstrated good size-exclusion effect for the adsorption of standard protein lysozyme with molecular weight (MW) of ca. 15 kDa; and the peptides with MW lower than this value can be well adsorbed. Step elution of the enriched peptides with five salt concentrations presented that both modified nanoparticles have high capacity and complementarity for peptides enrichment, and the SAX-MCM-41 nanoparticles has obviously high selectivity for acidic peptides with pI (isoelectric point) lower than 4. Large-scale enrichment of endogenous peptides in 2 mg mouse liver extract was achieved by further combination of SCX-MCM-41 and SAX-MCM-41 with unmodified MCM-41 nanoparticles. On-line 2D nano-LC/MS/MS was applied to analyze the enriched samples, and 2721 unique peptides were identified in total. Two-dimensional analysis of MW versus pI distribution combined with abundance of the identified peptides demonstrated that the three types of nanoparticles have comprehensive complementarity for peptidome enrichment. (c) 2008 Published by Elsevier B.V.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据