▎ 摘　　要

In this work, a novel size-exclusive metal oxide affinity chromatography (SE-MOAC) platform was built for phosphoproteome research. The operation for preparing graphene @titania @mesoporous silica nanohybrids (denoted as G@TiO2@mSiO(2)) was facile and easy to conduct by grafting titania nanoparticles on polydopamine (PD)-covered graphene, following a layer of mesoporous silica was coated on the outermost layer. The G@TiO2@mSiO(2) nanohybrids exhibited high sensitivity with a low detection limit of 5 amol/mu L (a total amount of 1 fmol) and high selectivity for phosphopeptides at a mass ratio of phosphopeptides to non-phosphopeptides (1:1000). The size-exclusive capability of the nanohybrids were also demonstrated by enriching the phosphopeptides from the mixture of Bovine Serum Albumin (BSA), alpha-casein, and beta-casein digests with a high mass ratio (beta-casein digests: alpha-casein: BSA, 1:500:500), which was attributed to the large surface area and ordered mesoporous channels. In addition, the G@TiO2@ mSiO(2) nanohybrids were employed to capture the endogenous phosphopeptides from human serum successfully. (C) 2015 Elsevier B.V. All rights reserved.