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Selective separation and enrichment of peptides for MS analysis using the microspheres composed of Fe3O4@nSiO2 core and perpendicularly aligned mesoporous SiO2 shell
Authors:Hemei Chen  Shasha Liu  Huili Yang  Yu Mao  Chunhui Deng  Xiangmin Zhang  Pengyuan Yang
Institution:1. Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, P. R. China;2. Center of Evaluation for Drug Safety, Second Military Medical University, Shanghai, P. R. China
Abstract:In this work, we report the development of a novel enrichment protocol for peptides by using the microspheres composed of Fe3O4@nSiO2 Core and perpendicularly aligned mesoporous SiO2 shell (designated Fe3O4@nSiO2@mSiO2). The Fe3O4@nSiO2@mSiO2 microspheres possess useful magnetic responsivity which makes the process of enrichment fast and convenient. The highly ordered nanoscale pores (2 nm) and high‐surface areas of the microspheres were demonstrated to have good size‐exclusion effect for the adsorption of peptides. An increase of S/N ratio over 100 times could be achieved by using the microspheres to enrich a standard peptide, and the application of the microspheres to enrich universal peptides was performed by using myoglobin tryptic digest solution. The enrichment efficiency of re‐used Fe3O4@nSiO2@mSiO2 microspheres was also studied. Large‐scale enrichment of endogenous peptides in rat brain extract was achieved by the microspheres. Automated nano‐LC‐ESI‐MS/MS was applied to analyze the sample after enrichment, and 60 unique peptides were identified in total. The facile and low‐cost synthesis as well as the convenient and efficient enrichment process of the novel Fe3O4@nSiO2@mSiO2 microspheres makes it a promising candidate for selectively isolation and enrichment of endogenous peptides from complex biological samples.
Keywords:Automated nano‐LC‐ESI‐MS/MS  Endogenous peptides  Magnetic mesoporous microspheres  MALDI‐TOF MS  Nanoproteomics  Selective enrichment
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