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[rAAV-Edu] rAAV9の精製方法 – 特許,2018 – △ID2559 [2019/10/05]

[rAAV-Edu] rAAV9の精製方法 – 特許,2018 – △ID2559 [2019/10/05]

rAAV9 vectorの精製方法

培養液のバッファ組成をTFF処理で調整、硫安(AmSo4)沈殿処理と第4級アンモニウム陰イオン交換クロマトグラフィー(HiPrep Q XL)によるFlow through mode、最後にSECによる精製でrAAV9を得る

スポンサーリンク: ID6183593457

rAAV9 vectorの調製方法

  1. Transfection by 3 plasmid (with PEI, FBS-free)
  2. Post culture
  3. Harvest supernatant
  4. TFF process
  5. 1/3 sut. concentration AmSO4 precipitation
  6. centrifuge
    • to take the sup.
  7. 1/2 sut. concentration AmSO4 precipitation
  8. centrifuge
    • to take the sup.
  9. Dilution to 7.3 mS/cm
  10. AEX
    • HiPress Q XL 16/10 column chromatograph with flow through mode
  11. TFF process
    • 30kDa
  12. SEC column chromatography
    • HiLoad 16/60 Superdex 200, MNH pH6.5, 300mM NaCl, 0.01% F-68

Highly Efficient Ultracentrifugation-free
Chromatographic Purification of Recombinant
AAV Serotype 9

https://www.cell.com/molecular-therapy-family/methods/pdfExtended/S2329-0501(18)30111-6

Improved methods for purification of recombinant aav vectors, EP2443233A1, 2009

https://patents.google.com/patent/EP2443233A1/en

用語の解説リンク、関連投稿および広告

AAV

[用語] rAAV [Biotech] [2022/09/03]

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AAV9

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rAAV-production

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スポンサーリンク ID 7130582967

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