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Highly Efficient Ultracentrifugation-free Chromatographic Purification of Recombinant AAV Serotype 9
http://hdl.handle.net/2241/00154784
http://hdl.handle.net/2241/00154784c3ac2813-4a6f-47da-b85d-3d54c17a7b98
名前 / ファイル | ライセンス | アクション |
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MTMCD_11 (1.7 MB)
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Item type | Journal Article(1) | |||||
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公開日 | 2019-03-06 | |||||
タイトル | ||||||
タイトル | Highly Efficient Ultracentrifugation-free Chromatographic Purification of Recombinant AAV Serotype 9 | |||||
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 |
石井, 亜紀子
× 石井, 亜紀子× 玉岡, 晃× Tomono, Taro× Hirai, Yukihiko× Okada, Hironori× Miyagawa, Yoshitaka× Adachi, Kumi× Sakamoto, Shuhei× Kawano, Yasuhiro× Chono, Hideto× Mineno, Junichi× Shimada, Takashi× Onodera, Masafumi× Okada, Takashi |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Recombinant adeno-associated virus serotype 9 (rAAV9) can specifically transduce muscle and neuronal tissues; thus, rAAV9 can potentially be used in gene therapy. However, rAAV9 is the most challenging rAAV serotype to purify. Traditionally, rAAV9 has been purified by ultracentrifugation, which is not scalable. We recently described a chromatographic purification protocol for rAAV1; this protocol can achieve scalable purifications. In this study, we attempted to optimize this protocol for purifying rAAV9 preparations, and we developed a novel, effective method for high-yield purification of rAAV9 using quaternary ammonium anion exchangers and size-exclusion chromatography. The final purified rAAV9 contained mainly three capsid proteins, as observed by SDS-PAGE. Furthermore, negative-stain electron microscopy demonstrated that 96.1% ± 1.1% of rAAV9 particles carried the viral genome containing the EGFP transgene, indicating that impurities and empty capsids can be eliminated with our purification protocol. The final rAAV9 titer obtained by our protocol totaled 2.5 ± 0.4 × 1015 viral genomes produced from ∼3.2 × 109 HEK293EB cells. We confirmed that our protocol can also be applied to purify other varied AAV genome constructs. Our protocol can scale up production of pure rAAV9, in compliance with current good manufacturing practice, for clinical applications in human gene therapy. | |||||
書誌情報 |
Molecular Therapy - Methods & Clinical Development 巻 11, p. 180-190, 発行日 2018-12 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 23290501 | |||||
PubMed番号 | ||||||
識別子タイプ | PMID | |||||
関連識別子 | 30533449 | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1016/j.omtm.2018.10.015 | |||||
権利 | ||||||
権利情報 | © 2018 The Authors. | |||||
権利 | ||||||
権利情報 | This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). | |||||
著者版フラグ | ||||||
値 | publisher | |||||
出版者 | ||||||
出版者 | American Society of Gene & Cell Therapy | |||||
出版者 | ||||||
出版者 | Elsevier |