蛋白质组学与生物信息学杂志
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开发用于固定胰蛋白酶以快速蛋白质消化的核壳磁性介孔SiO2微球

齐大伟、邓永辉、刘颖超、林华清、邓春辉、李艳、张向民、杨鹏远、赵东远

在工作中，我们开发了缩水甘油氧基丙基三甲氧基硅烷(GLYMO)-modified Fe3O4@SiO2 core and perpendicularly aligned mesoporous SiO2 shell (designated Fe3O4@nSiO2@mSiO2) as the novel substrate for the immobilization of large amount of trypsin and applied it for fast protein digestion. Firstly, Fe3O4@nSiO2@mSiO2 microspheres were synthesized. Then, the surface of the microspheres was functionalized with GLYMO for enzyme immobilization.The amount of trypsin immobilized on Fe3O4@nSiO2@mSiO2 was about 188 ? g/mg, which was much more than that on the previous magnetic materials. Using the trypsin-immobilized magnetic mesoporous SiO2 microspheres, proteins in samples were fast digested with microwave irradiation. The efficacy of this technique for protein mapping was demonstrated by the mass spectral analysis of the peptide fragmentation of three standard proteins, including cytochrome c (Cyt-c), myglobin (MYG), and bovine serum albumin (BSA). The functionalized magnetic microspheres served not only as substrate for enzyme immobilization, but also as excellent microwave absorbers, thus greatly improved the efficiency of protein digestion. It is also worth noting that by using this novel approach, the protein can be effectively digested within seconds, in contrast to hours required by conventional methods. Moreover, the trypsin-immobilized magnetic mesoporous SiO2 microspheres exhibit better stability than conventional methods. Furthermore, the feasibility of using this novel strategy for real sample analysis was demonstrated by applying it to the analysis of human垂体提取为其在大规模蛋白质组分析中的进一步应用开辟了途径。