摘要
构建了以阳离子交换色谱-反相色谱(SCX-RPLC)为分离模式的新型全二维微柱液相色谱-质谱分离平台。采用了醋酸铵缓冲液梯度洗脱,实现了第一维肽段的分步洗脱,洗脱的肽段经富集除盐后通过接口进入反相色谱微柱,通过线性梯度实现第二维进一步分离,最后进入质谱进行检测。采用此平台分析了人胃癌组织与正常组织提取蛋白质信息,其中正常胃组织鉴定蛋白质数为537个,而癌症组织鉴定蛋白质数目为506个。对胃癌和正常组织两种提取蛋白质酶解产物的蛋白质检索结果进行比较分析,将鉴定的蛋白质按照物理性质进行分布,找出正常组织与癌症组织间蛋白质差异,筛选出一种可能发生变异的癌症特有蛋白。
A two-dimensional high performance liquid chromatographic separation platform based on strong cation exchange（SCX） chromatography and reversed phase（RP） chromatography was developed.Peptides were first separated by a SCX column under stepwise gradient of 0,20,200,300,500,1000 and 2000 mmol/L NH4AC buffer.After each fraction was collected and desalted by a RP trap column,further separation was implemented on a RPLC column under linear gradient elution.All the components were identified by mass spectrometry after 2D separation.The platform was applied to the analysis of proteins in stomach cancer tissue.The number of identified proteins were 537 and 506 for healthy and tumor tissue respectively.According to the physical characters of proteins identified,the distributions of these identified proteins were obtained and the difference between healthy and tumor tissue was found.The retrieval results were compared to screen out diagnostic proteins related to pathological changes of tumor tissues.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2009年第10期1403-1408,共6页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金重点基金(No.20435020)
国家自然科学基金面上基金(No.20675083)
"973"计划(No.2007CB914102)资助项目
关键词
胃癌组织
蛋白质组
二维液相色谱-质谱联用
Stomach cancer tissue, proteomics, two-dimensional high performance liquid chromatographymass spectrometry
