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论文类型：期刊论文

发表时间：2014-05-01

发表刊物：TALANTA

收录刊物：SCIE、EI、PubMed、Scopus

卷号：122

页面范围：278-284

ISSN号：0039-9140

关键字：Graphene oxide; Dendrimer; On-plate digestion; Proteomics; IMER

摘要：In this study, dendrimer grafted graphene oxide nanosheets (dGO) were prepared by covalent reaction. The successful synthesis of dGO was confirmed by Fourier-transform infrared spectra, Raman spectra, Thermo gravimetric analysis and Zeta potential. Taking advantages of large surface area, excellent biocompatibility and abundant functional groups, dGO provided an ideal substrate for trypsin immobilization. Trypsin-linked dGO was synthesized through covalent bonding using glutaraldehyde as coupling agents. The amount of trypsin immobilized on dGO nanosheets was calculated to be about 649 + 20 mg/ g. The activity of immobilized trypsin could be maintained for over 10 days at 4 C. On-plate proteolysis could be performed without removing trypsin-linked dGO, because dGO did not interfere with matrixassisted laser desorption ionization time-of-flight tandem mass spectrometry analysis. By such an immobilized enzymatic reactor, standard proteins could be efficiently digested within 15 min, with sequence coverages comparable or better than those obtained by conventional over-night in-solution digestion. Furthermore, trypsin-linked dGO showed high sensitivity when applied to trace samples analysis. All these results demonstrated that the developed dGO based enzymatic reactor might provide a promising tool for high throughput proteome identification. (C) 2014 Elsevier B.V. All rights reserved.