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学科主题: 分析化学
题名: An integrated platform for quantitative proteome analysis with combination of protein labeling, separation and peptide separation via online digestion
作者: Zhou Y(周愿) ;  Yuan HM(袁辉明) ;  Tao DY(陶定银) ;  Zhang LH(张丽华) ;  Zhang YK(张玉奎)
会议文集: Human Proteome Organisation
会议名称: Human Proteome Organisation
会议日期: 2011-9-4
出版日期: 2011
会议地点: 日内瓦
通讯作者: 张丽华
出版者: 待补充
出版地: 待补充
合作性质: 墙报
部门归属: 1810
主办者: 波恩大学
摘要: At present, the hyphenation of isotopic labeling and mass spectrometry (MS) techniques is one of the most dominant approaches in quantitative proteomics, for example, traditional “shotgun” method coupled with stable isotope labeling of proteins or peptides has been widely used for quantitative proteome analysis [1]. However, it still suffered from some unavoidable drawbacks such as poor reproducibility, pushing the researchers to develop alternative approaches. In this report, an integrated platform [2] with the combination of protein labeling by stable isotope, separation by a mixed-bed WAX/WCX column, online digestion by a hydrophilic immobilized enzymatic reactor and peptide separation by high reproducible HPLC-chip was established. To evaluate the performance of such an integrated platform, Bovine serum albumin labeled by formaldehyde and formaldehyde-D2 with the ratio of 1:1 was analyzed, and the results demonstrated that the ratio of light and heavy dimethyl labels was 0.9998 and the RSD of the peptides identified from four runs was less than 10%. Furthermore, a mixture of bovine serum albumin, cytochrome C and myoglobin labeled by CH2O and CD2O with the ratio of 1:1 was also investigated and the median of peak intensity of all peptide was 0.9914. All these results demonstrated that such an integrated platform would be a promising alternative to traditional quantitative proteomic approach with high accuracy and high analysis throughput.
英文摘要: At present, the hyphenation of isotopic labeling and mass spectrometry (MS) techniques is one of the most dominant approaches in quantitative proteomics, for example, traditional “shotgun” method coupled with stable isotope labeling of proteins or peptides has been widely used for quantitative proteome analysis [1]. However, it still suffered from some unavoidable drawbacks such as poor reproducibility, pushing the researchers to develop alternative approaches. In this report, an integrated platform [2] with the combination of protein labeling by stable isotope, separation by a mixed-bed WAX/WCX column, online digestion by a hydrophilic immobilized enzymatic reactor and peptide separation by high reproducible HPLC-chip was established. To evaluate the performance of such an integrated platform, Bovine serum albumin labeled by formaldehyde and formaldehyde-D2 with the ratio of 1:1 was analyzed, and the results demonstrated that the ratio of light and heavy dimethyl labels was 0.9998 and the RSD of the peptides identified from four runs was less than 10%. Furthermore, a mixture of bovine serum albumin, cytochrome C and myoglobin labeled by CH2O and CD2O with the ratio of 1:1 was also investigated and the median of peak intensity of all peptide was 0.9914. All these results demonstrated that such an integrated platform would be a promising alternative to traditional quantitative proteomic approach with high accuracy and high analysis throughput.
语种: 英语
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/116019
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:
Zhou Y,Yuan HM,Tao DY,et al. An integrated platform for quantitative proteome analysis with combination of protein labeling, separation and peptide separation via online digestion[C]. 见:Human Proteome Organisation. 日内瓦. 2011-9-4.
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