摘要
以TiO_2介孔气凝胶作为富集材料,考察了其对标准糖蛋白辣根过氧化物酶(HRP)酶解物的富集效果。结果表明,流动相中乙腈浓度和pH对富集结果均有影响,以高有机相酸性条件上样清洗,低有机相碱性条件洗脱为佳。其次以牛血清白蛋白(BSA)为干扰蛋白,考察了TiO_2介孔气凝胶材料对糖肽的富集特异性,并与商品TiO_2颗粒的富集效果进行了对比。结果表明,TiO_2介孔气凝胶材料可在10倍的BSA干扰下实现HRP的选择性富集,富集能力和富集选择性显著优于商品化TiO_2颗粒,这归因于TiO_2介孔气凝胶材料较高的比表面积。研究有助于更好的理解TiO_2介孔气凝胶材料的保留机理,并将其应用于实际样品的预处理中。
Selective glycopeptide enrichment is one of the most important steps in glycoproteome research. In this work mesoporous TiO2 aerogel matrix was applied to selectively enrich glyeopeptides from tryptic digest of horseradish peroxidase （HRP）. Primarily, the influence of mobile phase to enrichment selectivity was evaluated, and both acetonitrile content and pH in the mobile phase could influence the glycopeptide enrichment selectivity. The optimized sample loading and eluting conditions were high acetonitrile content with acid added and low acetonitrile content with basic added, respectively. Secondly, the disturbed experiments were carried out to test the glycopeptide enrichment effect of the method by using the mixture of HRP and BSA （ molar ratio of 1 ： 10 ） as the target sample. The enrichment capability and selectivity of mesoporous TiO2 aerogel matrix were remarkably higher than that of commercial TiO2 particles, which was attributed to the high specific surface area. The investigation helps the researchers better understand the merit of mesoporous TiO2 aerogel matrix and better apply the matrix to real biosamples.
出处
《分析试验室》
CAS
CSCD
北大核心
2017年第5期604-608,共5页
Chinese Journal of Analysis Laboratory
关键词
氧化钛介孔气凝胶
糖肽
选择性富集
辣根过氧化物酶
Mesoporous TiO2 aerogel matrix
Glycopeptide
Selective enrichment
Horseradish peroxidase
