DICP OpenIR
学科主题物理化学
Multidimensional liquid chromatography based integrated platform for proteome study
Yuan HM(袁辉明); Ma JF(马俊锋); Zhou Y(周愿); Sun LL(孙良亮); Liang Z(梁振); Zhang LH(张丽华); Zhang YK(张玉奎)
会议名称Post-genome Technologies(IFPT6)
会议日期2009-9-17
2009-09-17
会议地点中国
页码381/1
部门归属十八室
主办者清华大学,东南大学,南京大学,西北大学
英文摘要To achieve high efficiency, high resolution, high accuracy and high throughput analysis of proteomes, an on-line integrated platform with the combination of proteins separation by 2D-SEC-RPLC, removal of organic modifier by an hollow fiber dialysis membrane, protein digestion by immobilized enzymatic reactor (IMER), peptides trapping by parallel C18 trapping columns, peptides separation by RPLC, and protein identification by ESI-MS/MS, was established, and applied for the analysis of proteins extracted from mouse brain. In our protocol, proteins were first separated by SEC with an analytical column. With a time interval of 10 min, eluants were divided into three fractions, and in turn trapped and desalted on a C8 trapping column, followed by separation by RPLC. For each RPLC run, the eluant was further divided into three fractions, with the time interval of 100 min. Before on-line digestion with IMER, each fraction from (RPLC was passed through a hollow fiber membrane based interface (with cut of molecular weight of 3 000), to on-line remove acetonitrile (ACN). To fasten such a procedure, 50 mM NH4 HCO3 containing 5% (v/v) CAN was used as the exchange buffer, with the flow rate of 2 mL/min. Subsequently, proteins could be on-line digested by IMER with trypsin immobilized on hybrid monolith inorganic-organic matrix without stop flow. Finally, peptides were separated and identified by RPLC-ESI-MS/MS. With such a platform, 1 055 proteins could be identified from mouse brain within 18 hr. All these results demonstrate that such an integrated platform might be promising to achieve high throughput analysis of proteomes with improved resolution, accuracy and throughput.
语种中文
文献类型会议论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/113632
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
推荐引用方式
GB/T 7714
Yuan HM,Ma JF,Zhou Y,et al. Multidimensional liquid chromatography based integrated platform for proteome study[C],2009:381/1.
条目包含的文件
条目无相关文件。
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[袁辉明]的文章
[马俊锋]的文章
[周愿]的文章
百度学术
百度学术中相似的文章
[袁辉明]的文章
[马俊锋]的文章
[周愿]的文章
必应学术
必应学术中相似的文章
[袁辉明]的文章
[马俊锋]的文章
[周愿]的文章
相关权益政策
暂无数据
收藏/分享
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。