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学科主题: 物理化学
题名: Integrated capillary electrophoresis platform for protein analysis
作者: Wang TT(王婷婷) ;  Ma JF(马俊锋) ;  Zhu GJ(朱贵杰) ;  Liang Z(梁振) ;  Zhang LH(张丽华) ;  Zhang YK(张玉奎)
会议名称: The 24th MSB 2009
会议日期: 2009-10-19
出版日期: 2009-10-19
会议地点: 中国
通讯作者: 张丽华
部门归属: 十八室
主办者: 大连化物所
摘要: For bottom-up strategy, proteins are digested into peptides before analyzed by HPLC-MS/MS. However, the large number of peptides not only brings tremendous challenge to chromatography separation, but also might result in false positive identifications. Therefore, intact proteins separation before digestion might be helpful. Recently, Dovichi et al integrated the intact proteins separation by CSE, on-line digestion by pepsin immobilized microreactor, the separation of resulting peptides by MEKC and the protein identified by MS/MS, and showed the promising of the combination of top-down and bottom-up strategies [1]. In our recent work, a novel integrated capillary electrophoresis platform, combining CIEF with monolithic immobilized pH gradient (M-IPG) materials for protein separation, trypsin immobilized microenzymatic reactor for on-line protein digestion, and CZE for peptide separation was established, and successfully applied for protein analysis. In the integrated platform, a tee unit was used to connect M-IPG column and microreactor, and to supply buffer to improve the compatibility of protein separation and digestion. Another interface was made by Teflon tube to improve the compatibility of microreactor and CZE. By such a platform, a 3-protein-mixture was analyzed, and 15 fractions from M-IPG CIEF were in sequence transferred to microreactor, and further separated by CZE. With collected peptides identified by MALDI-TOF MS, 4-peptide of myoglobin and 7-peptide of BSA were identified by such an integrated M-IPG CIEF-microreactor-CZE platform, which shows the potential of such a platform in high throughput protein analysis.
语种: 中文
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/113618
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:
Wang TT,Ma JF,Zhu GJ,et al. Integrated capillary electrophoresis platform for protein analysis[C]. 见:The 24th MSB 2009. 中国. 2009-10-19.
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