caa a22 4500 445852542 CHVBK 20180317145417.0 cr unu---uuuuu 170323e20110101xx s 000 0 eng 10.1007/s00204-010-0539-3 doi (NATIONALLICENCE)springer-10.1007/s00204-010-0539-3 Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform [Elektronische Daten] [Peng Zhao, Lijuan Zhang, Weijian Zhong, Xianping Ying, He Huang, Biyun Yao, Zhun Yuan, Juanling Fu, Yuedan Wang, Zongcan Zhou] 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-9,10-epoxide-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. Springer-Verlag, 2010 Antigen subtraction nationallicence Two-dimensional gel electrophoresis nationallicence Q-TOF MS/MS nationallicence anti-Benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (anti-BPDE) nationallicence anti-BPDE-transformed cells nationallicence Zhao Peng Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Zhang Lijuan Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Zhong Weijian Department of Toxicology, Shanghai Municipal Center for Disease Control & Prevention, 200336, Shanghai, People's Republic of China aut Ying Xianping Department of Toxicology, Shanghai Municipal Center for Disease Control & Prevention, 200336, Shanghai, People's Republic of China aut Huang He Department of Microbiology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Yao Biyun Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Yuan Zhun Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Fu Juanling Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Wang Yuedan Department of Immunology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Zhou Zongcan Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut Archives of Toxicology Springer-Verlag 85/1(2011-01-01), 35-41 0340-5761 85:1<35 2011 85 204 https://doi.org/10.1007/s00204-010-0539-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00204-010-0539-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhao Peng Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Lijuan Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhong Weijian Department of Toxicology, Shanghai Municipal Center for Disease Control & Prevention, 200336, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Ying Xianping Department of Toxicology, Shanghai Municipal Center for Disease Control & Prevention, 200336, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Huang He Department of Microbiology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Yao Biyun Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Yuan Zhun Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Fu Juanling Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Wang Yuedan Department of Immunology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhou Zongcan Department of Toxicology, Peking University Health Science Center, 100191, Beijing, People's Republic of China aut NATIONALLICENCE 773 0- Archives of Toxicology Springer-Verlag 85/1(2011-01-01), 35-41 0340-5761 85:1<35 2011 85 204 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer