DICP OpenIR
Subject Area分析化学
An integrated platform for quantitative proteome analysis with combination of protein labeling, separation and peptide separation via online digestion
Zhou Y(周愿); Yuan HM(袁辉明); Tao DY(陶定银); Zhang LH(张丽华); Zhang YK(张玉奎)
Source PublicationHuman Proteome Organisation
Conference NameHuman Proteome Organisation
Conference Date2011-9-4
2011
Conference Place日内瓦
Pages153-0
Publisher待补充
Publication Place待补充
Cooperation Status墙报
Department1810
Funding Organization波恩大学
AbstractAt 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.; 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.
Language英语
Document Type会议论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/116019
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang LH(张丽华)
Recommended Citation
GB/T 7714
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]. 待补充:待补充,2011:153-0.
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