基于反相有机整体柱的新型蛋白质酶反应器

摘要

以牛血清白蛋白(BSA)为标准样品,以鉴定的独立肽段数目、蛋白质鉴定覆盖率和漏切率为考察指标,通过考察酶解缓冲液、反相柱基质的多孔结构、酶解反应时间和整体柱的疏水性对反相蛋白质酶反应器酶解效率的影响,构建了一个基于反相有机整体柱的新型酶反应器,此反应器具有制备简单、通透性好、酶解时间短和蛋白质鉴定覆盖率高等特点.%X We developed a reversed phase monolithic column based enzyme reactor for protein analysis. Using the number of identified unique peptides, sequence coverage and the level of missed cleavage as the evaluation indicators, we systematically investigated the influence of buffer solution, porous structure of the column, digestion time and hydrophobicity of solid phase on the digestion performance of enzyme reactor. It had demonstrated that the reversed phase monolithic column based enzyme reactor had high digestion efficiency and might be further extended to the proteome analysis of complex samples.