摘要
本研究制备了一种应用于液-质联用(LC-MS)系统中的带喷头混合型毛细管色谱柱.用标准蛋白和酵母蛋白胰蛋白酶酶切溶液考察了其在LC-MS中的离子化效率、毛细管色谱柱分离性能和寿命,并与直接填充型毛细管色谱柱的色谱性能进行比较,未见显著性差异.将制备的带喷头混合型毛细管色谱柱应用于鼠肝蛋白组的分析,在假阳性率为1%的条件下鉴定到1262个肽段,归属于513个蛋白簇.通过对鉴定蛋白质理化性能的统计分析,其等电点和分子量分布高于一般两维凝胶电泳的范围.实验结果还表明该混合型毛细管色谱柱在肽段的理化性能上没有偏性,可以广泛应用于蛋白质组学的研究中.
A capillary chromatographic column with an emitter at one end is prepared with monolithic material in the emitter as a cribellum for the capillary column by using lauryl methacrylate-ethylene dimethacrylate （LMA-EDMA）, and then the other empty part of the capillary column with an emitter is packed with octadecylsilica particles（ODS）. The property of monolithic part of the capillary column is similar to that of reversed-phase packing materials packed in the capillary column and benefits the improvement on separation efficiency and prevents the clogging problem of the capillary column in the tapered tip during its use. The performances of the packing material-mixed-column such as peak capacity, ionization efficiency, signal intensity and the coverage of identified peptides or proteins are exam- ined on a nano-high performance liquid chromatography-mass spectrometry. We also evaluate the mixed-capillary column by analyzing trypticaUy digested proteins extracted from mouse livers. The results show that the capillary columns newly prepared possess a satisfied performance and will play an important role in proteomics researches in the future.
出处
《中国科学（B辑）》
CAS
CSCD
北大核心
2009年第8期752-759,共8页
Science in China(Series B)
基金
国家重点基础研究发展计划(编号:2006CB910803
2007CB914104)
国家高技术研究发展计划(编号:2006AA02A308)
国家自然科学基金(批准号:30621063
20635010
20735005
20875101)项目资助
关键词
蛋白质组学
多肽分离
毛细管色谱柱
液-质联用系统
proteomics, peptide separation, capillary chromatographic column, nano-high performance liquid chromatography-mass spectrometry
