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Ultracentrifugation-free chromatography-mediated large-scale purification of recombinant adeno-associated virus serotype 1 (rAAV1)
http://hdl.handle.net/2241/00154089
http://hdl.handle.net/2241/0015408992c84a2c-79d9-4cd5-abf1-b401aa467646
名前 / ファイル | ライセンス | アクション |
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MTMCD_3-2016-15058 (512.4 kB)
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Item type | Journal Article(1) | |||||
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公開日 | 2018-12-25 | |||||
タイトル | ||||||
タイトル | Ultracentrifugation-free chromatography-mediated large-scale purification of recombinant adeno-associated virus serotype 1 (rAAV1) | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源 | http://purl.org/coar/resource_type/c_6501 | |||||
タイプ | journal article | |||||
著者 |
Tomono, Taro
× Tomono, Taro× Hirai, Yukihiko× Okada, Hironori× Adachi, Kumi× Ishii, Akiko× Shimada, Takashi× Onodera, Masafumi× Tamaoka, Akira× Okada, Takashi |
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著者別名 |
石井, 亜紀子
× 石井, 亜紀子× 玉岡, 晃 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Recombinant adeno-associated virus (rAAV) is an attractive tool for gene transfer and shows potential for use in human gene therapies. The current methods for the production and purification of rAAV from the transfected cell lysate are mainly based on cesium chloride and iodixanol density ultracentrifugation, although those are not scalable. Meanwhile, chromatography-based systems are more scalable. Therefore, in this study, we developed a novel method for the production and purification of rAAV serotype 1 (rAAV1) from serum-free culture supernatant based on ion-exchange and gel-filtration chromatography to obtain highly purified products with an ultracentrifugation-free technique towards Good Manufacturing Practice (GMP) production. The purified rAAV1 displayed three clear and sharp bands (VP1, VP2, and VP3) following sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and more than 90% of rAAV1 particles contained fully packaged viral genomes according to negative-stain electron micrographic analysis. Consequently, the resultant genomic titer of the purified rAAV1 was 3.63 × 1013 v.g./ml (the total titer was 4.17 × 1013 v.g.) from the 4 × 109 HEK293 cells. This novel chromatography-based method will facilitate scale-up of manufacturing for clinical applications in gene therapy. | |||||
書誌情報 |
Molecular Therapy - Methods & Clinical Development 巻 3, p. 15058, 発行日 2016-12 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 2329-0501 | |||||
PubMed番号 | ||||||
識別子タイプ | PMID | |||||
関連識別子 | 26913289 | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1038/mtm.2015.58 | |||||
権利 | ||||||
権利情報 | © 2016 Official journal of the American Society of Gene & Cell Therapy. Published by Elsevier Inc. | |||||
権利 | ||||||
権利情報 | This work is licensed under a Creative Commons Attribution- NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/ licenses/by-nc-nd/4.0/ | |||||
著者版フラグ | ||||||
値 | publisher | |||||
出版者 | ||||||
出版者 | Elsevier |