Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1216, Issue 8, Pages 1270-1278Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2008.10.002
Keywords
Peptidomics; Ordered porous nanoparticles; Strong ion exchange; Reversed phase; Two-dimensional separation; Mouse liver
Funding
- 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
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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.
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