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学科主题: 物理化学
题名: Multidimensional liquid chromatography based integrated platform for high efficient analysis of intact proteins
作者: Yuan HM(袁辉明) ;  Zhang LH(张丽华) ;  Zhou Y(周愿) ;  Wu Q(吴琪) ;  Liang Z(梁振) ;  Zhang YK(张玉奎)
会议名称: 全球第九届人类蛋白质组学会议
会议日期: 2010-9-19
出版日期: 2010-09-19
会议地点: 澳大利亚
通讯作者: 张丽华
部门归属: 生物技术部
主办者: University of New South Wales
摘要: For proteome research, although MUDPIT and 2DPAGE are two mainstream technologies commonly used in most proteomics laboratories, they still suffer from many unavoidable drawbacks such as low efficiency, long analysis time and complex sample handling, pushing scientists to develop some alternative approaches. I n our recent study, a multidimensional integrated platform with the combination of protein separation by two-dimensional liquid chromatography, on-line solvent exchange by a hollow fiber membrane interface, on-line digestion by an immobilized enzymatic microreactor (IMER) as well as peptides separation and proteins identification by μRPLC/MS/MS was established. During the analysis, proteins were firstly separated by size exclusion chromatography (SEC). With a time interval of 15min, the eluants from SEC were divided into three fractions, and in turn trapped and desalted on a C8 precolumn, followed by μRPLC separation. For each μRPLC run, the eluants were further divided into three fractions with a time interval of 140 min, which was identical to that of peptide separation. After passing through a hollow fiber membrane interface to on-line remove organic modifier and adjust pH, each fraction from μRPLC was on-line digested by an IMER, and the resulting peptides were trapped on two parallel C18 precolumns. Finally, the peptides were separated and identified by μRPLC-ESI-MS/MS. With such a platform, more than 2000 proteins from rat brain were identified within 21 hr (FDR<1%). All these results demonstrated that our developed platform would be an attractive approach for high resolution and high throughput proteome analysis.
语种: 中文
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/114168
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Yuan HM,Zhang LH,Zhou Y,et al. Multidimensional liquid chromatography based integrated platform for high efficient analysis of intact proteins[C]. 见:全球第九届人类蛋白质组学会议. 澳大利亚. 2010-9-19.
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