改性介孔材料制备及其对低分子量蛋白质的在线选择富集研究

Investigation of the precolumn packed with modified silica-based mesoporous materials for online selective enrichment of low molecular weight proteins

制备了3种内外表面修饰不同官能团的硅基介孔材料,采用傅立叶变换红外光谱对修饰后材料的官能团进行了表征,并采用高效液相色谱对材料选择性富集低分子量蛋白质的能力进行了考查。3种材料对低分子量蛋白质和小肽均具有选择性富集能力,其中乙烯-二醇材料的富集能力最好。采用乙烯-二醇介孔材料装填预柱,考察了其对低分子量蛋白质的在线富集能力。高分子量蛋白质(卵清蛋白,分子量43000Da)在该预柱上没有保留,在死时间里直接流出柱外,而低分子量蛋白质(胰岛素,分子量5700Da)在该预柱上得到了较好的选择性萃取。保留在柱上的低分子量样品通过提高流动相中的有机相含量进行洗脱。研究工作对低分子量蛋白质和小肽的在线富集提供了一种新技术。

In this study,three mesoporous materials,alkyl-diol@ vinyl-Si O2,alkyl-diol@ methyl-Si O2 and alkyldiol@ methacryloxypropyl-Si O2 with modified exterior and interior surfaces were prepared by co-condensation and post-grafting and evaluated by using Fourier transform infrared spectra（ FTIR） and high performance liquid chromatography（ HPLC）. It was demonstrated that the new mesoporous materials all exhibited high selectivity for the extraction of low molecular weight（ LMW） proteins and peptides from protein mixtures,while alkyl-diol@vinyl-Si O2 showed the best. A precolumn was packed with alkyl-diol @ vinyl-Si O2 material and investigated by online extraction of the LMW protein（ insulin） with the interference of the high molecular weight（ HMW）protein（ ovalbumin）. The LMW proteins were selectively extracted on the precolumn,and eluted by the mobile phase containing a high portion of organic solvent.