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学科主题基础医学
Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform
Zhao, Peng2; Zhang, Lijuan2; Zhong, Weijian3; Ying, Xianping3; Huang, He4; Yao, Biyun2; Yuan, Zhun2; Fu, Juanling2; Wang, Yuedan1; Zhou, Zongcan2
关键词Antigen subtraction Two-dimensional gel electrophoresis Q-TOF MS/MS anti-Benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (anti-BPDE) anti-BPDE-transformed cells
刊名ARCHIVES OF TOXICOLOGY
2011
DOI10.1007/s00204-010-0539-3
85期:1页:35-41
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Toxicology
研究领域[WOS]Toxicology
关键词[WOS]MANGANESE CHLORIDE ; CELLS
英文摘要

In the present study, we developed a novel proteomic research strategy named antigen-subtracted 2-DE/MS strategy and applied it to comparative proteomic analysis of anti-benzo(a)pyrene-7,8-dihydrodiol-9,10epoxide-transformed human bronchial epithelial cell line (16HBE-C) and its parental cell line (16HBE) G0/G1 cells. Following pre-purification by ammonium sulfate precipitation, rabbit antibodies against 16HBE G0/G1 cells were coupled with protein A/G PLUS-agarose under the maximal coupling rate of about 50%. The agarose-antibody complex was then used in immunoprecipitation known as antigen subtraction. When the mass ratio of antibody to the sample was 2.5-3:1, the subtraction rates for 16HBE and 16HBE-C G0/G1 cell proteins were 44 and 34%, respectively. Both subtracted and unsubtracted samples were then subjected to the 2-DE resolution. In 16HBE-subtracted 2-DE maps, 315 protein spots were subtracted and 49 new protein spots were detected, whereas in 16HBE-C-subtracted 2-DE maps, 287 protein spots were subtracted and 33 new protein spots were detected. Taken together, antigen subtraction results in 65 new protein spots being allowed to be detected, therefore, makes it possible to get more information of the samples. Finally, 4 protein spots only detected in 16HBE-C-subtracted 2-DE maps were analyzed by the Q-TOF MS/MS, and successfully identified as U6 snRNA-associated Sm-like protein LSm3, 60S acidic ribosomal protein P1, Peroxiredoxin-6 and 60S acidic ribosomal protein P2. These proteins are involved in carcinogenesis, oxidation stress and protein synthesis. In conclusion, the antigen-subtracted 2-DE/MS strategy could reduce the complexities of protein samples and therefore, improve the resolution for the sample analysis.

语种英语
WOS记录号WOS:000286620900006
项目编号2002CB512906 ; 30471470 ; 30671781 ; 30771995
资助机构China National Key Basic Research and Development Program ; National Nature Science Foundation of China
引用统计
被引频次:4[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/56833
专题北京大学基础医学院_免疫学系
北京大学医学部管理机构_人事处
北京大学基础医学院
北京大学公共卫生学院_公共卫生学院
作者单位1.Peking Univ, Hlth Sci Ctr, Dept Immunol, Beijing 100191, Peoples R China
2.Peking Univ, Hlth Sci Ctr, Dept Toxicol, Beijing 100191, Peoples R China
3.Shanghai Municipal Ctr Dis Control & Prevent, Dept Toxicol, Shanghai 200336, Peoples R China
4.Peking Univ, Hlth Sci Ctr, Dept Microbiol, Beijing 100191, Peoples R China
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GB/T 7714
Zhao, Peng,Zhang, Lijuan,Zhong, Weijian,et al. Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform[J]. ARCHIVES OF TOXICOLOGY,2011,85(1):35-41.
APA Zhao, Peng.,Zhang, Lijuan.,Zhong, Weijian.,Ying, Xianping.,Huang, He.,...&Zhou, Zongcan.(2011).Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform.ARCHIVES OF TOXICOLOGY,85(1),35-41.
MLA Zhao, Peng,et al."Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform".ARCHIVES OF TOXICOLOGY 85.1(2011):35-41.
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