杂化整体材料在芯片-质谱系统中富集糖蛋白的研究

Bio-mass spectrometry plays an important role in the characterization of proteins. When bio-mass spectrometry is applied to the analysis of glycoproteins, it often suffers from the disadvantages of low ionization of glycosylated peptides. In addition, the ion signals of glycosylated peptides in mass spectrometry analysis are always suppressed by the large amount of non-glycosylated peptides. The goal of the present study is to develop an effective and selective method for the enrichment of glycoproteins. Nanometer Al2O3 will be prepared and employed for the modification of organic polymer monolith. The prepared inorganic-organic hybrid monolithic material in a microchip will be used for the determination of glycoproteins. The enrichment capacities of glycosylated and non-glycosylated peptides with the inorganic-organic hybrid monolithic material will be studied in detail. Combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) as the detector, a microchip platform will thus be successfully established for the analysis of glycosylated peptides. With the present strategy, a novel and effective method may be provided to enhance the sensitivities and selectivities for the determination of glycoproteins with mass spectrometry.

生物质谱是解析蛋白质肽段结构的重要手段。在质谱分析糖基化蛋白质过程中，糖基化肽段离子化效率较低，不易被质谱识别鉴定，且质谱信号常常受到非糖基化肽段的严重抑制和干扰。本项目针对蛋白质组学研究中面临的糖基化蛋白质高效选择性富集方面的难点问题，拟制备纳米氧化铝并将其用于有机聚合物整体柱的改性，在微流控芯片系统内制备无机-有机杂化整体材料，以期实现糖基化蛋白质的高效富集。研究无机-有机整体材料对糖基化和非糖基化肽段的选择性吸附富集能力；利用基质辅助激光解析质谱(MALDI-MS)对富集后的糖基化肽段进行分析鉴定，在此基础上建立适用于糖基化蛋白质分析的微流控芯片平台。本项目的实施将为提高糖基化蛋白质质谱检测的灵敏度、选择性提供有效的研究方法，推进质谱方法在糖基化蛋白质分析中的应用进程。

整体柱已逐渐发展成微流控芯片分析系统中最佳的预处理模式。将无机材料与聚合物整体柱杂化制备无机-有机杂化整体柱，其中的无机组份可提供机械强度稳定的聚合骨架，而结合于无机相上均匀分布的有机基团可改善材料的性能，在分离选择性研究方面显示了特殊的优势。本项目成功实现了γ-Al2O3等纳米粒子包覆的一系列聚合物整体柱的制备，由于无机纳米粒子是由化学键引入聚合物整体柱孔表面的，该整体柱具有良好的通透性、稳定的机械性能和高的萃取容量。本项目的实施为提高分析检测灵敏度、选择性提供了有效的研究方法，项目执行期间发表SCI收录论文20篇。