新型游离胶分馏器OFFGEL对SH-SY5Y细胞蛋白酶切后肽段分离性能考察  

Peptide Fractionation of the Digested SH-SY5Y Proteins by OFFGEL

作　　者：张永谦[1] 才德[1] 段晋燕[1] 王洪彬[1] 邓玉林[1] 

机构地区：[1]北京理工大学生命学院,北京100081

出　　处：《化学通报》2010年第9期837-841,共5页Chemistry

基　　金：国家"十一.五"科技支撑计划项目(2006BAK03A07);北京理工大学研究生科技创新基金项目(AA200805)资助

摘　　要：在基于鸟枪法鉴定蛋白的策略中,需要在肽段水平上进行高效良好的预分离。以人多巴胺能神经细胞瘤(SH-SY5Y)为研究对象,考察了游离胶分馏器(OFFGEL)分离肽段的性能。SH-SY5Y细胞全酶切肽段经过OFFGEL分离后,得到24个不同馏分,每一个馏分经过微流液相色谱-质谱检测,发现有83.7%的肽段是专属于一个馏分中,各个馏分中肽段平均等电点与理论值基本一致,且在pH低端肽段的等电点偏差较小,达到良好的聚焦效果。OFFGEL不但能同时分离多达16个样品,而且能够从溶液中回收样品,具有高通量、操作简单、方便后续质谱检测等优点。因此,OFFGEL在蛋白质组学中多肽的分离具有很好的应用前景。Efficient separation in the peptide level is crucial to the protein identification based on the shotgun strategy.The ability of peptide fractionation of OFFGEL was investigated by tryptic digested peptides of SH-SY5Y.24 fractions were obtained after OFFGEL fractionation.Each fraction was performed by capillary high performance liquid chromatography electrospray mass spectrometry.83.7% peptides were unique to one fraction and average isoelectric point values in each fraction were basically consistent with the expected values.Furthermore,the standard derivations ofisoelectric point values were small in the low pH range.OFFGEL could fractionated 16 samples simultaneously,recovery the sample from solution directly and facilitate to the detection by mass spectrometry.OFFGEL can play more and more important role in the peptide separation.

关 键 词：游离胶分馏器 蛋白质组学 液相色谱质谱 

分 类 号：Q51[生物学—生物化学]