Proteomics analysis of co-purifying cellular proteins associated with rAAV vectors.
Proteomics analysis of co-purifying cellular proteins associated with rAAV vectors.
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DOI：
10.1371/journal.pone.0086453
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Xiao W
中科院分区：
文献类型：
--
作者：
Dong B;Duan X;Chow HY;Chen L;Lu H;Wu W;Hauck B;Wright F;Kapranov P;Xiao W
Recombinant adeno-associated vectors (rAAV) are commonly purified by either chromatography or equilibrium CsCl gradient. Nevertheless, even after purification various cellular proteins often associate with rAAV vector capsids. Such co-purifying cellular proteins may raise concern about safety of gene therapy. Here we report identification and characterization of the co-purifying cellular protein in the vector preparations by using a combination of two proteomics approaches, GeLC-MS (gel electrophoresis liquid chromatography-mass spectrometry) and 2DE (two-dimensional gel electrophoresis). Most prominent bands revealed by Coomassie Blue staining were mostly similar to the AAV capsid proteins. Posttranslational modifications of capsid proteins were detected by the proteomics analysis. A total of 13 cellular proteins were identified in the rAAV vectors purified by two rounds of cesium chloride gradient centrifugation, including 9 by the GeLC-MS analysis and 4 by the 2DE analysis. Selected cellular proteins were verified by western blot. Furthermore, the cellular proteins could be consistently found associated with different AAV serotypes and carrying different transgenes. Yet, the proteins were not integral components of the viral capsis since a stringent washing procedure by column purification could remove them. These co-purified proteins in AAV vector preparations may have a role in various stages of the AAV life cycle.
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DOI：
10.1073/pnas.0702778104
发表时间：
2007-08-07
影响因子：
11.1
作者：
Wang, Jinhui;Xie, Jing;Xiao, Weidong
通讯作者：
Xiao, Weidong
影响因子：
4.4
作者：
Duan, Xunbao;Kelsen, Steven G.;Merali, Salim
通讯作者：
Merali, Salim
影响因子：
--
作者：
Gray, Steven J;Choi, Vivian W;Asokan, Aravind;Haberman, Rebecca A;McCown, Thomas J;Samulski, Richard Jude
通讯作者：
Samulski, Richard Jude
影响因子：
3.4
作者：
Duan, Xunbao;Kelsen, Steve G.;Merali, Salim
通讯作者：
Merali, Salim
影响因子：
5.5
作者：
Aslanidi, George V.;Rivers, Angela E.;Ortiz, Luis;Govindasamy, Lakshmanan;Ling, Chen;Jayandharan, Giridhara R.;Zolotukhin, Sergei;Agbandje-McKenna, Mavis;Srivastava, Arun
通讯作者：
Srivastava, Arun