Purification of infectious adenovirus in two hours by ultracentrifugation and tangential flow filtration
文献类型：期刊论文
| 作者 | Ugai, H; Yamasaki, T; Hirose, M; Inabe, K; Kujime, Y; Terashima, M; Liu, BB; Tang, H; Zhao, MJ; Murata, T |
| 刊名 | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
 |
| 出版日期 | 2005 |
| 卷号 | 331期号:4页码:1053-1060 |
| 关键词 | adenovirus recombinant adenovirus purification ultracentrifugation tangential flow filtration |
| 通讯作者 | Yokoyama, KK (reprint author), RIKEN, Inst Phys & Chem Res, Tsukuba Inst, Gene Engn Div,BioResource Ctr, 3-1-1 Koyadai, Tsukuba, Ibaraki 3050074, Japan.,kazu@brc.riken.jp |
| 英文摘要 | Adenoviruses are excellent vectors for gene transfer and are used extensively for high-level expression of the products of transgenes in living cells. The development of simple and rapid methods for the purification of stable infectious recombinant adenoviruses (rAds) remains a challenge. We report here a method for the purification of infectious adenovirus type 5 (Ad5) that involves ultracentrifugation on a cesium chloride gradient at 604,000g for 15 min at 4 degrees C and tangential flow filtration. The entire procedure requires less than two hours and infectious Ad5 can be recovered at levels higher than 64% of the number of plaque-forming units (pfu) in the initial crude preparation of viruses. We have obtained titers of infectious purified Ad5 of 1.35 x 10(10) pfu/ml and a ratio of particle titer to infectious titer of seven. The method described here allows the rapid purification of rAds for studies of gene function in vivo and in vitro, as well as the rapid purification of Ad5. (c) 2005 Elsevier Inc. All rights reserved. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | ANION-EXCHANGE HPLC ; RECOMBINANT ADENOVIRUS ; GENE-THERAPY ; IMMUNE-RESPONSE ; VECTORS ; PARTICLES ; PROTEIN ; CELLS ; P53 ; ACTIVATION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000229234500024 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1892]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Ugai, H,Yamasaki, T,Hirose, M,et al. Purification of infectious adenovirus in two hours by ultracentrifugation and tangential flow filtration[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2005,331(4):1053-1060. |
| APA | Ugai, H.,Yamasaki, T.,Hirose, M.,Inabe, K.,Kujime, Y.,...&Yokoyama, KK.(2005).Purification of infectious adenovirus in two hours by ultracentrifugation and tangential flow filtration.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,331(4),1053-1060. |
| MLA | Ugai, H,et al."Purification of infectious adenovirus in two hours by ultracentrifugation and tangential flow filtration".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 331.4(2005):1053-1060. |
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