聚酰基氨基脲类多孔有机聚合物的制备及其在糖肽富集中的应用

摘要

通过均苯三甲酰肼(BTZ)和对苯二异氰酸酯(PDI)的聚合反应制备了一种新型的聚酰基氨基脲类多孔有机聚合物(POP).采用傅里叶变换红外光谱(FT-IR)、交叉极化魔角自旋碳13核磁共振(CP-MAS 13C NMR)、氮气物理吸附/脱附实验和水接触角等手段对POP-1的物理性质进行表征.其中比表面积为166 m2/g,水接触角为46.4°,表明这种多孔材料具有良好的亲水性.因此,尝试将POP-1应用于标准蛋白质和复杂生物样品中糖肽的亲水色谱富集.通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/MS)分析,从500 fmol人血清免疫球蛋白G(IgG)酶解液中共鉴定到19条典型的N-连接糖肽;通过毛细管液相色谱-串联质谱(cLC-MS/MS)分析,从100 μg鼠肝蛋白酶解液中共鉴定到属于605个糖蛋白的1919个糖肽的1350个N-糖基化位点.这些结果表明POP-1在糖蛋白质组学研究中具有较强的应用潜力.%An acylsemicarbazide-based porous organic polymer (POP) was facilely prepared by the polymerization of benzene-1,3,5-tricarbohydrazide (BTZ) and 1,4-phenylene diisocyanate (PDI).The physical properties of as-synthesized material were characterized by Fourier-transform infrared spectroscopy (FT-IR),solid-state cross polarisation magic angle spinning carbon-13 nuclear magnetic resonance (CP-MAS 13C NMR),nitrogen adsorption/desorption measurement,water contact angle and so on.The specific surface area was 166 m2/g,and the water contact angle was 46.4°,exhibiting hydrophilic property of the porous material.Thus,POP-1 was tried to be used for glycopeptides enrichment from tryptic digest of standard protein and complex biosamples in hydrophilic mode.The 19 typical N-linked glycopeptides were identified from 500 fmol tryptic digest of immunoglobulin G (IgG) from human serum by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis,meanwhile,1919 unique glycopeptides with 1350 N-glycosylation sites from 605 different N-linked glycoproteins were identified from 100 μg mouse liver tryptic digest by capillary liquid chromatography (cLC)-MS/MS analysis.These results demonstrated the great potential of POP-1 for glycoproteome analysis.