[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
- Transfection by 3 plasmid (with PEI, FBS-free)
- Post culture
- Harvest supernatant
- TFF process
- 1/3 sut. concentration AmSO4 precipitation
- centrifuge
- to take the sup.
- 1/2 sut. concentration AmSO4 precipitation
- centrifuge
- to take the sup.
- Dilution to 7.3 mS/cm
- AEX
- HiPress Q XL 16/10 column chromatograph with flow through mode
- TFF process
- 30kDa
- SEC column chromatography
- HiLoad 16/60 Superdex 200, MNH pH6.5, 300mM NaCl, 0.01% F-68
Highly Efficient Ultracentrifugation-free
https://www.cell.com/molecular-therapy-family/methods/pdfExtended/S2329-0501(18)30111-6
Chromatographic Purification of Recombinant
AAV Serotype 9
Improved methods for purification of recombinant aav vectors, EP2443233A1, 2009
https://patents.google.com/patent/EP2443233A1/en